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Information on EC 7.2.1.1 - NADH:ubiquinone reductase (Na+-transporting) and Organism(s) Vibrio cholerae serotype O1 and UniProt Accession A5F5Y6
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7.2.1.1 NADH:ubiquinone reductase (Na+-transporting)IUBMB Comments
An iron-sulfur flavoprotein, containing two covalently bound molecules of FMN, one noncovalently bound FAD, one riboflavin, and one [2Fe-2S] cluster.
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Word Map
- 7.2.1.1
- vibrio
- cholerae
- flavin
- fmn
- harveyi
- medicine
-
redox-driven
- riboflavin
- alginolyticus
-
one-electron
- ubiquinol
-
flavosemiquinone
-
na+-dependent
-
phosphoester
-
flavinylation
- hqno
- 2-n-heptyl-4-hydroxyquinoline
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
na+-nqr, na(+)-translocating nadh:quinone oxidoreductase, na+-translocating nadh:quinone oxidoreductase, na(+)-translocating nadh:ubiquinone oxidoreductase, nqr-1, nqr-2, na(+)-pumping nadh:quinone oxidoreductase, na+-translocating nadh:ubiquinone oxidoreductase, sodium-translocating nadh:quinone oxidoreductase, na+-translocating nadh-quinone reductase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
dNADH:K3 oxidoreductase
-
Na(+)-translocating NADH-quinone reductase subunit A
-
Na(+)-translocating NADH-quinone reductase subunit B
Na(+)-translocating NADH-quinone reductase subunit D
-
Na(+)-translocating NADH-quinone reductase subunit E
-
Na(+)-translocating NADH:ubiquinone oxidoreductase
-
Na+-NQR
Na+-pumping NADH: quinone oxidoreductase
-
Na+-pumping NADH:quinone oxidoreductase
Na+-translocating NADH dehydrogenase
-
Na+-translocating NADH:quinone oxidoreductase
Na+-translocating NADH:ubiquinone oxidoreductase
NADH:quinone oxidoreductase
-
NQR-1
complex which elutes at an approximate molecular mass of 306 kDa is termed NQR-1. One copy of each Nqr subunit is present in the Na+-NQR (NqrA-His, NqrF, NqrB, NqrC, NqrD and NqrE). NQR-1 mainly consists of monomeric Na+-NQR
NQR-2
the smaller subcomplex of the Na+-NQR consists of NqrA-His, NqrF, NqrB and NqrCNQR-2 but lacks the highly hydrophobic NqrD and NqrE subunits
NqrB
-
sodium ion-translocating NADH:quinone oxidoreductase
sodium-translocating NADH:quinone oxidoreductase
Na(+)-translocating NADH-quinone reductase subunit B
-
Na(+)-translocating NADH-quinone reductase subunit B
-
Na+-NQR
-
-
Na+-NQR
-
Na+-NQR
-
Na+-pumping NADH:quinone oxidoreductase
-
-
Na+-pumping NADH:quinone oxidoreductase
-
Na+-pumping NADH:quinone oxidoreductase
-
Na+-translocating NADH:quinone oxidoreductase
-
-
Na+-translocating NADH:quinone oxidoreductase
-
Na+-translocating NADH:quinone oxidoreductase
-
Na+-translocating NADH:ubiquinone oxidoreductase
-
sodium ion-translocating NADH:quinone oxidoreductase
-
SYSTEMATIC NAME
IUBMB Comments
NADH:ubiquinone oxidoreductase (Na+-translocating)
An iron-sulfur flavoprotein, containing two covalently bound molecules of FMN, one noncovalently bound FAD, one riboflavin, and one [2Fe-2S] cluster.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 ferricyanide + NADH + n Na+/in
2 ferrocyanide + NAD+ + H+ + n Na+/out
-
-
-
?
deamino-NADH + H+ + ubiquinone + 2 Na+/in
deamino-NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + 2 Na+/in
NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + n Li+/in
NAD+ + ubiquinol + n Li+/out
-
-
-
-
?
NADH + H+ + ubiquinone + Na+/in
NAD+ + ubiquinol + Na+/out
-
-
-
?
NADH + H+ + ubiquinone 1
NAD+ + ubiquinol 1
-
-
-
?
NADH + H+ + ubiquinone-1 + n Li+/in
NAD+ + ubiquinol-1 + n Li+/out
-
-
-
?
NADH + H+ + ubiquinone-8 + n Na+/in
NAD+ + ubiquinol-8 + n Na+/out
-
-
-
?
reduced nicotinamide hypoxanthine dinucleotide + menadione
oxidized nicotinamide hypoxanthine dinucleotide + menadiol
-
-
-
?
thio-NADH + H+ + ubiquinone + 2 Na+/in
thio-NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
-
the Na+-pumping NADH:ubiquinone oxidoreductase is a molecular energy transducer present in the membrane of many marine and pathogenic bacteria. The enzyme accepts electrons from NADH and donates them to ubiquinone. This redox reaction releases a significant amount of energy, which is used to pump Na+ across the cell membrane, creating a Na+ gradient as well as an electrical potential. Electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
Vibrio cholerae and many other marine and pathogenic bacteria possess a unique respiratory complex, the Na+-pumping NADH:quinone oxidoreductase, which pumps Na+ across the cell membrane using the energy released by the redox reaction between NADH and ubiquinone
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
-
electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
-
the results on the reverse reaction, in which the oxidized enzyme takes electrons from quinol, confirm that riboflavin, the center that gives rise to the neutral flavosemiquinone, is located downstream of both FMNB and FMNC in the forward pathway. Riboflavin is likely to be the site from which electrons are donated to quinone during enzyme turnover
-
-
r
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
-
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
NqrB-G140 and -G141 are critical for the binding and reaction of Na+-NQR with its electron acceptor, ubiquinone
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
the electrons move through the different redox centers in the enzyme in a linear pathway. During the first step of electron transfer, the non-covalently bound FAD accepts two electrons from NADH. Subsequently, the electrons are transferred stepwise by passing to the 2Fe-2S center, the two FMN molecules covalently attached to NqrC and NqrB (in this order), riboflavin, and finally to ubiquinone-8. The one-electron reduction of FMN in NqrC is the step involved in sodium uptake, and the reduction of riboflavin is involved in sodium translocation
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
-
the large, peripheral NqrA subunit of the Na+-NQR binds one molecule of ubiquinone-8 with high affinity, which is determined by the methoxy groups at the C-2 and C-3 positions of the quinone headgroup. Ubiquinone-8 bound to NqrA occupies a functional site. Analysis of dynamic interaction of NqrA with quinones by surface plasmon resonance and saturation transfer difference NMR. Electron transfer in Na+-NQR is initiated by NADH oxidation on subunit NqrF and leads to quinol formation on subunit NqrA
-
-
?
NADH + H+ + ubiquinone-1
NAD+ + ubiquinol-1
-
-
-
?
NADH + H+ + ubiquinone-1
NAD+ + ubiquinol-1
the enzyme catalyzes NADH-driven Na+ transport
-
-
?
NADH + H+ + ubiquinone-1
NAD+ + ubiquinol-1
-
the NqrF subunit catalyzes the initial oxidation of NADH
-
-
?
NADH + H+ + ubiquinone-1 + n Na+/in
NAD+ + ubiquinol-1 + n Na+/out
-
-
-
?
NADH + H+ + ubiquinone-1 + n Na+/in
NAD+ + ubiquinol-1 + n Na+/out
-
-
-
?
NADH + H+ + ubiquinone-1 + n Na+/in
NAD+ + ubiquinol-1 + n Na+/out
-
subunit NqrA of the Na+-NQR harbours a Q binding site, and the catalytic site of quinone reduction is located on NqrA
-
-
?
NADH + H+ + ubiquinone-1 + n Na+/in
NAD+ + ubiquinol-1 + n Na+/out
ubiquinone-1 acts as substrate for the oxidation of NADH by the holo-Na+-NQR coupled to Na+ translocation. Ubiquinoone-1 also accepts electrons in a non-physiological reaction catalyzed by isolated NqrF or subcomplexes containing the NqrF subunit. Both the coupled and the non-physiological modes of NADH oxidation with Q1 require FAD as cofactor
-
-
?
NADH + ubiquinone-1
NAD+ + ubiquinol-1
-
-
-
?
additional information
?
-
-
the enzyme is the entry point for electrons into the respiratory chain
-
-
?
additional information
?
-
-
the NADH oxidation is coupled to the pumping of Na+ across the membrane
-
-
?
additional information
?
-
-
in addition to the quinone reductase reaction the isolated enzyme can also catalyze so-called NADH dehydrogenase reaction during interaction with soluble quinones. This activity includes a single-electron reduction of soluble quinones (menadione, Q0, Q1, etc.) or some other electron acceptors (hexammineruthenium(III), ferricyanide, etc.). Similarly to the transdehydrogenase activity, the NADH dehydrogenase activity does not depend on concentration of sodium ions, is inhibited by heavy metal ions, and is insensitive to 2-heptyl-4-hydroxyquinoline N-oxide and korormicin
-
-
?
additional information
?
-
-
in addition to the quinone reductase reaction, the isolated enzyme can also catalyze the so-called NADH-dehydrogenase reaction during interaction with soluble quinones. This activity includes a single-electron reduction of soluble quinones (menadione, Q0, Q1, etc.) or some other electron acceptors (hexaammineruthenium (III), ferricyanide, etc.)
-
-
?
additional information
?
-
-
Na+-NQR does not oxidize NADPH
-
-
?
additional information
?
-
-
2,5-dibromo-3-methyl-6-isopropyl-4-benzoquinone both acts as an inhibitor and as an alternative substrate of the Na+-NQR of Vibrio cholerae by a specific interaction with the NqrA subunit of the complex
-
-
?
additional information
?
-
-
binding activity of subunit NqrA with different quinones, i.e. quinone, ubiquinone-1, ubiquinone-2, ubiquinone-8, 3-azido-2-methoxy-5-methyl-6-geranyl-1,4-benzoquinone, 2-azido-3-methoxy-5-methyl-6-geranyl-1,4-benzoquinone, 3-azido-2-methoxy-5-methyl-6-geranyl-1,4-benzoquinone-biotin, biotinylated 3-azido-2-methoxy-5-methyl-6-geranyl-1,4-benzoquinone, overview
-
-
?
additional information
?
-
-
Na+-NQR enables pumping of Li+, as well as Na+ across the membrane, but the enzyme is not able to translocate other monovalent cations, such as potassium or rubidium
-
-
?
additional information
?
-
proposed mechanism. Redox-induced conformational changes critically couple electron transfer to Na+ translocation from the cytoplasm to the periplasm through a channel in subunit NqrB
-
-
?
additional information
?
-
the coupling efficiency of NQR is influenced by the nature of the transported cation, and by the concentration of protons. Partial uncoupling of the NQR observed with Li+, or with Na+ at pH 7.5-8.0, may be caused by the backflow of the coupling cation through the channel in subunit NqrB
-
-
?
additional information
?
-
the site of ubiquinone reduction is conformationally coupled to the NqrB subunit, which also hosts the final cofactor in the electron transport chain, riboflavin
-
-
?
additional information
?
-
the ubiquinone ring binds to the NqrA subunit in the regions Leu-32-Met-39 and Phe-131-Lys-138, encompassing the rear wall of a predicted ubiquinone-binding cavity. The binding sites for ubiquinone and aurachin-type inhibitors are in close proximity but do not overlap one another
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
deamino-NADH + H+ + ubiquinone + 2 Na+/in
deamino-NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + 2 Na+/in
NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + Na+/in
NAD+ + ubiquinol + Na+/out
-
-
-
?
NADH + H+ + ubiquinone-1
NAD+ + ubiquinol-1
the enzyme catalyzes NADH-driven Na+ transport
-
-
?
NADH + H+ + ubiquinone-1 + n Na+/in
NAD+ + ubiquinol-1 + n Na+/out
-
-
-
?
thio-NADH + H+ + ubiquinone + 2 Na+/in
thio-NAD+ + ubiquinol + 2 Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
-
the Na+-pumping NADH:ubiquinone oxidoreductase is a molecular energy transducer present in the membrane of many marine and pathogenic bacteria. The enzyme accepts electrons from NADH and donates them to ubiquinone. This redox reaction releases a significant amount of energy, which is used to pump Na+ across the cell membrane, creating a Na+ gradient as well as an electrical potential. Electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
-
-
?
NADH + H+ + ubiquinone
NAD+ + ubiquinol
Vibrio cholerae and many other marine and pathogenic bacteria possess a unique respiratory complex, the Na+-pumping NADH:quinone oxidoreductase, which pumps Na+ across the cell membrane using the energy released by the redox reaction between NADH and ubiquinone
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
-
-
-
-
?
NADH + H+ + ubiquinone + n Na+/in
NAD+ + ubiquinol + n Na+/out
-
-
-
?
additional information
?
-
-
the enzyme is the entry point for electrons into the respiratory chain
-
-
?
additional information
?
-
-
in addition to the quinone reductase reaction the isolated enzyme can also catalyze so-called NADH dehydrogenase reaction during interaction with soluble quinones. This activity includes a single-electron reduction of soluble quinones (menadione, Q0, Q1, etc.) or some other electron acceptors (hexammineruthenium(III), ferricyanide, etc.). Similarly to the transdehydrogenase activity, the NADH dehydrogenase activity does not depend on concentration of sodium ions, is inhibited by heavy metal ions, and is insensitive to 2-heptyl-4-hydroxyquinoline N-oxide and korormicin
-
-
?
additional information
?
-
-
in addition to the quinone reductase reaction, the isolated enzyme can also catalyze the so-called NADH-dehydrogenase reaction during interaction with soluble quinones. This activity includes a single-electron reduction of soluble quinones (menadione, Q0, Q1, etc.) or some other electron acceptors (hexaammineruthenium (III), ferricyanide, etc.)
-
-
?
additional information
?
-
-
Na+-NQR does not oxidize NADPH
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
putative arrangement of subunits and cofactors of the Na+-NQR
-
FAD
-
contains one molecule of covalently bound FAD, believed to be bound to subunit F
FAD
contains one noncovalently bound FAD
FAD
the electron transfer NADH -> FAD ->[2Fe-2S] in NqrF requires positioning of the FAD and the Fe-S cluster in close proximity in accordance with a structural model of the subunit
FAD
-
electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
FAD
the isolated enzyme complex contains near-stoichiometric amounts of non-covalently bound FAD (0.78 mol/mol Na+-NQR) and riboflavin (0.70 mol/mol Na+-NQR). The four flavins in the Na+-NQR are converted to the partially or fully reduced state when the enzyme reacts with NADH. Two non-covalently bound flavins (FAD and riboflavin) preferentially act as two-electron mediators, whereas two covalently bound FMNs undergo one-electron transitions (FMN/FMNU(radical)-)
FAD
-
the NqrF subunit contains one FAD and a [2Fe2S] cluster and catalyzes the initial oxidation of NADH
FAD
non-covalently bound in the NqrF subunit
FAD
noncovalently bound FAD
FAD
complex contains one non-covalently bound FAD, one noncovalently bound riboflavin, ubiquinone-8 and a [2Fe2S] cluster. The phosphate group is attached at the 5'-position of the ribityl as in authentic FMN and the NADH:quinone oxidoreductase contains approximately 1.7 mol covalently bound FMN per mol non-covalently bound FAD
FAD
the reduced FAD cofactor of cytoplasmic NqrF subunit is the site for intracellular superoxide formation
flavin
-
contains a single extremely stable flavin semiquinone, which becomes deprotonated upon reduction of the enzyme
FMN
contains two covalently bound FMNs
FMN
-
the enzyme contains two molecules of covalently bound FMN, covalently bound to subunits NqrB and NqrC
FMN
-
electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
FMN
four flavins in the Na+-NQR are converted to the partially or fully reduced state when the enzyme reacts with NADH. Two non-covalently bound flavins (FAD and riboflavin) preferentially act as two-electron mediators, whereas two covalently bound FMNs undergo one-electron transitions (FMN/FMNU(radical)-)
FMN
-
contains two FMNs covalently bound to the NqrB and NqrC subunits
FMN
subunits NqrB and NqrC contain covalently bound FMN, the wild type subunit NqrC contains approximately 1 mol of FMN per mol of protein
FMN
covalently bound FMN
FMN
two covalently bound in subunits NqrB und NqrC
NADH
-
-
NADH
-
NADH
-
NADH
NQR-2 exhibits approximately 15% of the specific NADH dehydrogenase activity of NQR-1
riboflavin
-
riboflavin
contains one riboflavin
riboflavin
-
electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
riboflavin
riboflavin detected in NQR-2 is bound to the NqrB or NqrC subunit
riboflavin
-
riboflavin is an active redox cofactor of Na+-NQR, giving rise to the neutral flavosemiquinone, it is likely to be the final electron carrier in the enzyme before the quinone
riboflavin
the isolated enzyme complex contains near-stoichiometric amounts of non-covalently bound FAD (0.78 mol/mol Na+-NQR) and riboflavin (0.70 mol/mol Na+-NQR). The four flavins in the Na+-NQR are converted to the partially or fully reduced state when the enzyme reacts with NADH. Two non-covalently bound flavins (FAD and riboflavin) preferentially act as two-electron mediators, whereas two covalently bound FMNs undergo one-electron transitions (FMN/FMNU(radical)-)
riboflavin
-
the enzyme contains one noncovalently bound riboflavin
riboflavin
noncovalently bound riboflavin in purified Na+-NQR is not liberated from FMN but represents an intrinsic component of the Na+-NQR complex
riboflavin
one non-covalently bound in the subunit NqrB
riboflavin
complex contains one non-covalently bound FAD, one noncovalently bound riboflavin, ubiquinone-8 and a [2Fe2S] cluster.. The phosphate group is attached at the 5'-position of the ribityl as in authentic FMN and the NADH:quinone oxidoreductase contains approximately 1.7 mol covalently bound FMN per mol non-covalently bound FAD. Each of the single NqrB and NqrC subunits in the NADH:quinone oxidoreductase carries a single FMN
ubiquinone-8
complex contains one non-covalently bound FAD, one noncovalently bound riboflavin, ubiquinone-8 and a [2Fe2S] cluster
[2Fe-2S]-center
contains one 2Fe-2S-center
[2Fe-2S]-center
the NqrF subunit of the Na+-NQR complex harbors a [2Fe-2S] cluster which accepts electrons from the FAD cofactor
[2Fe-2S]-center
complex contains one non-covalently bound FAD, one noncovalently bound riboflavin, ubiquinone-8 and a [2Fe2S] cluster
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe
Fe2+
Iron
K+
Li+
Na+
[2Fe-2S]
Fe
-
enzyme contains a 2Fe-2S center. Electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
Fe
the enzyme contains 650-780 ng Fe per mg protein, which corresponds to 2.5-3.0 atoms of Fe per enzyme complex. The enzyme bears only the 2Fe-2S cluster as the sole metal-containing prosthetic group
Fe
-
the enzyme contains one 2Fe-2S center, electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
Fe
-
the NqrF subunit contains one FAD and a [2Fe2S] cluster and catalyzes the initial oxidation of NADH. The Na+-NQR is proposed to increase the stability of NqrF' by stimulating the maturation of FeS centers
Fe2+
2 [2Fe-2S] clusters in the NqrF subunit
Iron
contains one 2Fe-2S center. Four conserved cysteines, C70, C76, C79, and C111 in the subunit NqrF match the canonical 2Fe-2S motif
K+
-
acts as a nonessential activator, increasing the activity and affinity for sodium. Na+-NQR contains a regulatory site for K+
K+
Na+-NQR undergoes significant conformational changes upon oxidoreduction, depending on the monovalent cation present (Na+, Li+, K+, or Rb+). Deprotonated acid residues are involved in the binding of cations, pointing toward a monodentate binding mode in the oxidized form of the enzyme and bidentate binding in the reduced form
Li+
the wild-type enzyme is stimulated 3fold by lithium, while the mutants NqrBD397A and NqrB-D397K, that are completely insensitive to sodium, are also not stimulated by lithium. The activities of the mutants NqrBD397C, NqrB-D397E, NqrB-D397N and NqrB-D397S are stimulated by lithium, and interestingly the fraction of stimulationis greater for lithium compared to sodium
Li+
in presence of Li+, transmembrane voltage is generally lower than in presence of Na+, and the pH profile of electron transfer activity does not reveal a pronounced maximum
Li+
Na+-NQR undergoes significant conformational changes upon oxidoreduction, depending on the monovalent cation present (Na+, Li+, K+, or Rb+). Deprotonated acid residues are involved in the binding of cations, pointing toward a monodentate binding mode in the oxidized form of the enzyme and bidentate binding in the reduced form. Residue D397 is one of the acid residues involved in Na+ and Li+ binding
Na+
-
recombinant enzyme has high specific activity in presence of Na+
Na+
environmental Na+ can stimulate ubisemiquinone formation by the Na+-NQR and hereby enhances the production of reactive oxygen species formed during the autoxidation of reduced quinines, increasing the Na+ concentration from below 0.1 mM to 25 mM results in a 3fold stimulation of activity of the purified enzyme
Na+
-
sodium ions are indispensable components of the Na+-NQR-catalyzed reaction, therefore its rate depends on concentration of Na+ and the reaction virtually does not occur in media depleted in this ion
Na+
-
specific requirement for sodium ions, even in the absence of sodium ions Na+-NQR is unable to translocate protons
Na+
the wild-type enzyme's physiologic activity is stimulated 8fold in the presence of saturating amounts of sodium
Na+
in presence of Na+, transmembrane voltage is barely influenced by pH (6.5-8.5), while quinone reduction activity exhibits a maximum at pH 7.5-8.0
Na+
Na+-NQR undergoes significant conformational changes upon oxidoreduction, depending on the monovalent cation present (Na+, Li+, K+, or Rb+). Deprotonated acid residues are involved in the binding of cations, pointing toward a monodentate binding mode in the oxidized form of the enzyme and bidentate binding in the reduced form. Residue D397 is one of the acid residues involved in Na+ and Li+ binding
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,5-dibromo-3-methyl-6-isopropyl-4-benzoquinone
-
a known inhibitor of the bc1 and b6f complexes found in mitochondria and chloroplasts, also inhibits quinone reduction by the Na+-NQR in a mixed inhibition mode. It does not just act as a simple competitor or redox mediator at the quinol oxidase site, but also as an antagonist to ubiquinone, inducing a redox bypass of the respiratory chain. The compound both acts as an inhibitor and as an alternative substrate of the Na+-NQR of Vibrio cholerae by a specific interaction with the NqrA subunit of the complex
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
-
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
-
aurachin D-42
the quinolone ring and alkyl side chain of inhibitor aurachin bind to the NqrB subunit in the regions Arg-43-Lys-54 and Trp-23-Gly-89, respectively
iodoacetamide
inactivation
Rb+
presence of Rb+ induces conformational changes, indicating a changed accessibility of the sodium binding sites
2-n-heptyl-4-hydroxyquinoline N-oxide
-
2-n-heptyl-4-hydroxyquinoline N-oxide
-
2-n-heptyl-4-hydroxyquinoline N-oxide
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
2-n-heptyl-4-hydroxyquinoline-N-oxide
-
Ag+
in the presence of Ag+, rates of NADH oxidation by membranes from the parent Vibrio cholera strain decrease to 14% (0.001 mM Ag+) of the activity observed in the absence of the inhibitor
Ag+
specific inhibition
korormicin
-
specifically inhibits Na+-NQR at the level of its interaction with ubiquinone
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
enzyme contains a 2Fe-2S center. Electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
FMN
-
enzyme contains a 2Fe-2S center. Electrons flow from NADH to quinone through the FAD in subunit F, the 2Fe-2S center, the FMN in subunit C, the FMN in subunit B, and finally riboflavin. The reduction of the FMN(C) to its anionic flavosemiquinone state is the first Na+-dependent process, suggesting that reduction of this site is linked to Na+ uptake. During the reduction reaction, two FMNs are transformed to their anionic flavosemiquinone in a single kinetic step. Subsequently, FMN(C) is converted to the flavohydroquinone, accounting for the single anionic flavosemiquinone radical in the fully reduced enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.4
n Li+/in
mutant D397N, pH 8.0, temperature not specified in the publication
2.5
n Li+/in
mutant D397E, pH 8.0, temperature not specified in the publication
2.7
n Li+/in
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
2.9
n Li+/in
mutant D397S, pH 8.0, temperature not specified in the publication
3.5
n Li+/in
wild-type, pH 8.0, temperature not specified in the publication
0.25
n Na+/in
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
0.7
n Na+/in
mutant D397C, pH 8.0, temperature not specified in the publication
1.8
n Na+/in
mutant D397E, pH 8.0, temperature not specified in the publication
2.5
n Na+/in
wild-type, pH 8.0, temperature not specified in the publication
2.8
n Na+/in
mutant D397S, pH 8.0, temperature not specified in the publication
3.1
n Na+/in
mutant D397N, pH 8.0, temperature not specified in the publication
0.0025
ubiquinone
wild-type enzyme, pH 8.0, temperature not specified in the publication
0.0088
ubiquinone
NqrB, mutant G141L, pH 8.0, temperature not specified in the publication
0.015
ubiquinone
NqrB, mutant G141A, pH 8.0, temperature not specified in the publication
0.0234
ubiquinone
NqrB, mutant G141V, pH 8.0, temperature not specified in the publication
0.1
ubiquinone
above, mutant NqrB-G140A, pH 8.0, temperature not specified in the publication
0.0025
ubiquinone-1
wild-type, pH 8.0, temperature not specified in the publication
0.0088
ubiquinone-1
NqrB, mutant G141L, pH 8.0, temperature not specified in the publication
0.015
ubiquinone-1
NqrB, mutant G141A, pH 8.0, temperature not specified in the publication
0.0234
ubiquinone-1
NqrB, mutant G141V, pH 8.0, temperature not specified in the publication
0.1
ubiquinone-1
mutant G140A, pH 8.0, temperature not specified in the publication
0.1
ubiquinone-1
mutant G140L, pH 8.0, temperature not specified in the publication
additional information
additional information
kinetics
-
additional information
additional information
Kmapp(Na+) is: 1.1 mM for wild-type enzyme, 1.4 mM for subunit D mutant D88L, 9.3 mM for subunit D mutant D133A
-
additional information
additional information
Kmapp(Na+) is: 1.1 mM for wild-type enzyme, 13.1 mM for subunit E mutant E95A
-
additional information
additional information
Kmapp(Na+) is: 1.1 mM for wild-type enzyme, 2.1 mM for subunit B mutant E28A, 4.2 mM for subunit B mutant E144L, 0.9 mM for subunit B mutant D346A, above 100 mM for subunit B mutant D397A
-
additional information
additional information
in wild-type Na+-NQR, the sodium dependence of the steady state turnover follows a Michaelis-Menten behavior, while the kinetics of the NqrB-D397A mutant show an unsaturable behavior for sodium. Kinetic mechanism, modeling, overview
-
additional information
additional information
-
Michaelis-Menten kinetics in the absence of 2,5-dibromo-3-methyl-6-isopropyl-4-benzoquinone
-
additional information
additional information
-
steady-state and transient kinetics, together with equilibrium binding measurements to define the number of cation-binding sites and characterize their roles in the enzyme, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
100
n Li+/in
mutant D397N, pH 8.0, temperature not specified in the publication
110
n Li+/in
mutant D397E, pH 8.0, temperature not specified in the publication
115
n Li+/in
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
124
n Li+/in
mutant D397S, pH 8.0, temperature not specified in the publication
180
n Li+/in
wild-type, pH 8.0, temperature not specified in the publication
115
n Na+/in
mutant D397S, pH 8.0, temperature not specified in the publication
127
n Na+/in
mutant D397E, pH 8.0, temperature not specified in the publication
127
n Na+/in
mutant D397N, pH 8.0, temperature not specified in the publication
170
n Na+/in
mutant D397C, pH 8.0, temperature not specified in the publication
170
n Na+/in
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
500
n Na+/in
wild-type, pH 8.0, temperature not specified in the publication
448.9
NADH
wild-type, pH 8.0, temperature not specified in the publication
487.7
NADH
mutant D397K, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
495.4
NADH
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
501.4
NADH
mutant D397C, pH 8.0, temperature not specified in the publication
511.7
NADH
mutant D397S, pH 8.0, temperature not specified in the publication
521.9
NADH
mutant D397K, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
544.7
NADH
mutant D397E, pH 8.0, temperature not specified in the publication
560.8
NADH
mutant D397N, pH 8.0, temperature not specified in the publication
580.2
NADH
mutant D397A, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
587.2
NADH
wild-type, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
600.1
NADH
wild-type, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
603.2
NADH
mutant D397S, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
622.2
NADH
mutant D397A, pH 8.0, temperature not specified in the publication
629.8
NADH
mutant D397A, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
656.6
NADH
mutant D397C, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
660.4
NADH
mutant D397N, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
662.9
NADH
mutant D397S, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
673.7
NADH
mutant D397E, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
681.6
NADH
mutant D397N, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
690.9
NADH
mutant D397C, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
694.6
NADH
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
695.4
NADH
mutant D397E, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
696.7
NADH
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
5199
NADH
mutant D397K, pH 8.0, temperature not specified in the publication
200
ubiquinone
mutant G140A, pH 8.0, temperature not specified in the publication
388
ubiquinone
mutant G141V, pH 8.0, temperature not specified in the publication
425
ubiquinone
mutant G141A, pH 8.0, temperature not specified in the publication
463
ubiquinone
mutant G141L, pH 8.0, temperature not specified in the publication
525
ubiquinone
wild-type enzyme, pH 8.0, temperature not specified in the publication
57.7
ubiquinone-1
mutant D397S, pH 8.0, temperature not specified in the publication
58.8
ubiquinone-1
mutant D397K, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
59.7
ubiquinone-1
mutant D397N, pH 8.0, temperature not specified in the publication
59.8
ubiquinone-1
mutant D397K, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
61.3
ubiquinone-1
mutant D397K, pH 8.0, temperature not specified in the publication
61.9
ubiquinone-1
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication
65.7
ubiquinone-1
mutant D397C, pH 8.0, temperature not specified in the publication
66
ubiquinone-1
mutant D397E, pH 8.0, temperature not specified in the publication
66.3
ubiquinone-1
wild-type, pH 8.0, temperature not specified in the publication
70.6
ubiquinone-1
mutant D397A, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
72.3
ubiquinone-1
mutant D397A, pH 8.0, temperature not specified in the publication
72.3
ubiquinone-1
mutant D397A, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
80.7
ubiquinone-1
mutant D397S, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
95.5
ubiquinone-1
mutant D397N, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
96.4
ubiquinone-1
mutant D397E, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
101.3
ubiquinone-1
mutant D397S, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
102.9
ubiquinone-1
mutant D397C, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
110.6
ubiquinone-1
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
122.9
ubiquinone-1
mutant D397N, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
129.6
ubiquinone-1
mutant D397E, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
152.9
ubiquinone-1
mutant D397C, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
162.2
ubiquinone-1
mutant D397C, presence of 2-mercaptoethanol, pH 8.0, temperature not specified in the publication, presence of 100 mM Na+
200
ubiquinone-1
mutant G140A, pH 8.0, temperature not specified in the publication
200
ubiquinone-1
mutant G140L, pH 8.0, temperature not specified in the publication
205.5
ubiquinone-1
wild-type, pH 8.0, temperature not specified in the publication, presence of 100 mM Li+
388
ubiquinone-1
mutant G141V, pH 8.0, temperature not specified in the publication
425
ubiquinone-1
mutant G141A, pH 8.0, temperature not specified in the publication
463
ubiquinone-1
mutant G141L, pH 8.0, temperature not specified in the publication
525
ubiquinone-1
wild-type, pH 8.0, temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
525
ubiquinone-1
wild-type, pH 8.0, temperature not specified in the publication
525
ubiquinone-1
mutant G140A, pH 8.0, temperature not specified in the publication
525
ubiquinone-1
mutant G140L, pH 8.0, temperature not specified in the publication
525
ubiquinone-1
mutant G141A, pH 8.0, temperature not specified in the publication
525
ubiquinone-1
mutant G141L, pH 8.0, temperature not specified in the publication
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
inhibition kinetics
-
0.0005
2-n-heptyl-4-hydroxyquinoline N-oxide
wild-type enzyme, pH 8.0, temperature not specified in the publication
0.0005
2-n-heptyl-4-hydroxyquinoline N-oxide
wild-type, pH 8.0, temperature not specified in the publication
0.0012
2-n-heptyl-4-hydroxyquinoline N-oxide
mutant G141V, pH 8.0, temperature not specified in the publication
0.0075
2-n-heptyl-4-hydroxyquinoline N-oxide
above, mutant G140A, pH 8.0, temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000013 - 0.01
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
0.00059 - 0.027
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
0.0001
Ag+
Vibrio cholerae serotype O1
pH and temperature not specified in the publication
0.0012
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
mutant G141A, pH 8.0, 30°C
0.002
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
wild-type, pH 8.0, 30°C
0.0031
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
mutant Y36A, pH 8.0, 30°C
0.0045
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
mutant G38V, pH 8.0, 30°C
0.2
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
mutant E144C, pH 8.0, 30°C
0.2
2-n-heptyl-4-hydroxyquinoline N-oxide
Vibrio cholerae serotype O1
mutant G140A, pH 8.0, 30°C
0.0000013
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
mutant G141A, pH 8.0, 30°C
0.0000016
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
wild-type, pH 8.0, 30°C
0.0000033
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
mutant Y36A, pH 8.0, 30°C
0.0000057
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
mutant G38V, pH 8.0, 30°C
0.00001
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
mutant E144C, pH 8.0, 30°C
0.01
3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
Vibrio cholerae serotype O1
mutant G140A, pH 8.0, 30°C
0.00059
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
wild-type, pH 8.0, 30°C
0.00083
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
mutant Y36A, pH 8.0, 30°C
0.00096
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
mutant G38V, pH 8.0, 30°C
0.0052
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
mutant G141A, pH 8.0, 30°C
0.027
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
mutant E144C, pH 8.0, 30°C
0.027
5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate
Vibrio cholerae serotype O1
mutant G140A, pH 8.0, 30°C
0.000002
aurachin D-42
Vibrio cholerae serotype O1
wild-type, pH 8.0, 30°C
0.0000027
aurachin D-42
Vibrio cholerae serotype O1
mutant Y36A, pH 8.0, 30°C
0.0000028
aurachin D-42
Vibrio cholerae serotype O1
mutant G38V, pH 8.0, 30°C
0.0000033
aurachin D-42
Vibrio cholerae serotype O1
mutant G141A, pH 8.0, 30°C
0.012
aurachin D-42
Vibrio cholerae serotype O1
mutant E144C, pH 8.0, 30°C
0.012
aurachin D-42
Vibrio cholerae serotype O1
mutant G140A, pH 8.0, 30°C
0.000005
korormicin
Vibrio cholerae serotype O1
wild-type, pH 8.0, 30°C
0.000008
korormicin
Vibrio cholerae serotype O1
mutant G38V, pH 8.0, 30°C
0.0000081
korormicin
Vibrio cholerae serotype O1
mutant Y36A, pH 8.0, 30°C
0.00082
korormicin
Vibrio cholerae serotype O1
mutant G141A, pH 8.0, 30°C
0.05
korormicin
Vibrio cholerae serotype O1
mutant E144C, pH 8.0, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
392715, 658968, 671141, 674705, 698842, 698995, 699385, 710746, 711269, 711293, 713414, 725433, 725448, 725538, 732791
-
-
A5F5X1: subunit NqrA, A5F5X0: subunit NqrB, A5F5Y7: subunit NqrC, A5F5Y6: subunit NqrD; A5F5Y3: subunit NqrE, A5F5Y4: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
UniProt
A5F5X1: subunit NqrA, A5F5X0: subunit NqrB, A5F5Y7: subunit NqrC, A5F5Y6: subunit NqrD; A5F5Y3: subunit NqrE, A5F5Y4: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
UniProt
A5F5X1: subunit NqrA, A5F5X0: subunit NqrB, A5F5Y7: subunit NqrC, A5F5Y6: subunit NqrD; A5F5Y3: subunit NqrE, A5F5Y4: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
UniProt
Q9KPS1: subunit NqrA, Q9KPS2: subunit NqrB, P0C6E0: subunit NqrC, Q9X4Q6: subunit NqrD, Q9X4Q7: subunit NqrE, Q9X4Q8: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
Uniprot
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
glycine 140 and glycine 141 of the NqrB subunit in the functional binding of ubiquinone. Mutations at these residues alter the affinity of the enzyme for ubiquinol. Mutations in residue NqrBG140 almost completely abolished the electron transfer to ubiquinone
metabolism
physiological function
additional information
metabolism
-
Na+-NQR couples the free energy of electron transfer reactions to electrogenic pumping of sodium across the cell membrane. Na+ uptake by Na+-NQR is a nonelectrogenic process that occurs upon electron transfer from the 2Fe-2S-center to FMN of subunit NqrC, while the electrogenic transport of Na+ across the membrane, and likely release into solution, occurs upon electron transfer from FMN of subunit NqrB to riboflavin
metabolism
the enzyme is the main entrance for electrons into the respiratory chain of many marine and pathogenic bacteria. The enzyme accepts electrons from NADH and donates them to ubiquinone, and the free energy released by this redox reaction is used to create an electrochemical gradient of sodium across the cell membrane
physiological function
-
the enzyme exploits the free energy liberated during oxidation of NADH with ubiquinone to pump sodium ions across the cytoplasmic membrane
physiological function
the enzyme is the entry site of electrons into the aerobic respiratory chain, catalyzing the electron transfer from NADH to ubiquinone, which is coupled to the pumping of sodium ions across the membrane. The sodium gradient produced by Na+-NQR is used by the cell for ATP synthesis, transport of nutrients, rotation of the flagellum, among other processes
physiological function
-
the Na+-translocating NADH:quinone oxidoreductase is the entry site for electrons into the respiratory chain and the main sodium pump in Vibrio cholerae
physiological function
the sodium ion-translocating NADH:quinone oxidoreductase from the human pathogen Vibrio cholerae is a respiratory membrane protein complex that couples the oxidation of NADHto the transport of Na+ across the bacterial membrane
physiological function
-
Vibrio cholerae relies on the Na+-pumping NADH:quinone oxidoreductase as the first complex in its respiratory chain
physiological function
-
the Vibrio cholerae Na+ translocating NADH:quinone oxidoreductase mutant shows multiple defects in metabolism. The mutant up-regulates 31 genes and down-regulates 55 genes in both early and mid-growth phases. The most up-regulated genes included the cadA and cadB genes, encoding a lysine decarboxylase and a lysine/cadaverine antiporter, respectively. The down-regulated genes include sialic acid catabolism genes. There is also an increase in the reductive pathway of TCA cycle and decreased purine metabolism in the mutant. Lack of Na+ translocating NADH:quinone oxidoreductase does not affect any of the Na+ pumping-related phenotypes of Vibrio cholerae
physiological function
In a deletion mutant devoid of the Na+-NQR encoding genes NqrA-NqrF, rates of respiratory Na+ extrusion are decreased by a factor of four, but the cytoplasmic Na+ concentration is essentially unchanged. The mutant is impaired in formation of transmembrane voltage and dioes not grow under hypoosmotic conditions at pH 8.2 or above. At alkaline pH and limiting Na+ concentrations, the Na+-NQR may be crucial for generation of a transmembrane voltage to drive the import of H+ by electrogenic Na+/H+ antiporters
physiological function
Na+-NQR acts as a as a producer of ROS in vivo. The amount of cytoplasmic ROS detected in Vibrio cholerae cells producing variants of Na+-NQR correlates well with rates of superoxide formation by the corresponding membrane fractions. Membranes from wild-type show increased superoxide production activity compared to membranes from a mutant lacking Na+-NQR. Overexpression of plasmid-encoded Na+-NQR results in a drastic increase in the formation of superoxide. The reduced FAD cofactor of cytoplasmic NqrF subunit is the site for intracellular superoxide formation
additional information
aspartate 397 in subunit B of the Na+-pumping NADH: quinone oxidoreductase forms part of a sodium binding site, is involved in cation selectivity and affects cation binding site cooperativity. The enzyme is partially functional with only one sodium-binding site, which indicates that the sites are connected to independent and probably parallel sodium pumping pathways
additional information
-
Na+-NQR is composed of six subunits NqrA-F and harbours at least five redox active cofactors: A non-covalently bound FAD and a 2Fe-2S cluster in the NqrF subunit, two covalently bound FMNs. The catalytic site of quinone reduction is located on NqrA. The two ligands bind to an extended binding pocket in direct vicinity to each other
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
110000
NQR-2 is the smaller subcomplex of the Na+-NQR consists of NqrA-His, NqrF, NqrB and NqrCNQR-2 but lacks the highly hydrophobic NqrD and NqrE subunit, gel filtration
213000
about, His6-tagged Na+-NQR complex, sequence calculation
21470
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
22837
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
27600
subunit NqrC, calculated from amino acid sequence
27619
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
306000
one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 5076 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, gel filtration
32000
recombinant subunit NqrC, SDS-PAGE
360000
45067
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
45357
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
48624
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
360000
-
it is likely that the protein is a monomer, calculated molecular mass of 215000 Da with bound detergent, gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
heterohexamer
hexamer
monomer
tetramer
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 45067 (Nqrf), NQR-2 is the smaller subcomplex of the Na+-NQR consists of NqrA-His, NqrF, NqrB and NqrCNQR-2 but lacks the highly hydrophobic NqrD and NqrE subunit, calculated from sequence
additional information
-
putative arrangement of subunits and cofactors of the Na+-NQR
heterohexamer
-
Na+-NQR is a heterohexameric complex consisting of six different protein subunits (NqrA-NqrF)
hexamer
the NqrF subunit harbors the active site of NADH oxidation and acts as a converter between the hydride donor NADH and subsequent one-electron reaction steps in the Na+-translocating NADH:quinone oxidoreductase complex
hexamer
1 * 48624 (NqrA) + 1 * 45357 (NqrB) + 1 * 27619 (NqrC) + 1 * 22837 (NqrD) + 1 * 21470 (NqrE) + 1 * 45067 (Nqrf), one copy of each Nqr subunit is present in the Na+-NQR complex and a theoretical molecular of 212000 Da is calculated for the complex. Taking into account the additional 50-76 kDa of a n-dodecyl beta-D-maltoside micelle which shields the hydrophobic part of the complex from the polar solvent, it is conluded that NQR-1 complex mainly consists of monomeric Na+-NQR, calculated from sequence
hexamer
-
Na+-NQR is a multisubunit, membrane-embedded NADH dehydrogenase
hexamer
-
the enzyme consists of four membrane-bound subunits NqrBCDE and the peripheral NqrF and NqrA subunits. The catalytic site of quinone reduction is located on subunit NqrA
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of theNa1-NQR complex at 3.5 A resolution. The electron transfer pathway starts from the NADH-oxidizing cytoplasmic NqrF subunit across the membrane to the periplasmic NqrC, and back to the quinone reduction site on NqrA located in the cytoplasm. A sodium channel is localized in subunit NqrB. In the mechanism of redox-driven Na+ translocation, the change in redox state of the flavin mononucleotide cofactor in NqrB triggers the transport of Na+ through the observed channel
sitting drop vapor diffusion method, using 40 mM KSCN, 21.0% PEG 2000 MME, 100 mM Tris acetate pH 8.5 and 8% (v/v) 1-propanol, cofactors such as FAD, riboflavin or NADH are not added during crystallization
-
structures of subunits NqrA, NqrC, and NqrF. An electron transfer pathway exists from cytoplasmic subunit NqrF across the membrane to the periplasmic subunit NqrC, and via subunit NqrB back to the quinone reduction site on cytoplasmic NqrA. A Fe site located in the midst of membrane-embedded subunits NqrD and NqrE shuttles the electrons over the membrane
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C111A
C112I
NqrD-C112I. The substitutions of the conserved cysteine residue in NqrD blocks the Na+-dependent and HQNO-sensitive quinone reductase activity of the enzyme, being without effect on the interaction of the enzyme with reduced nicotinamide hypoxanthine dinucleotide and menadione. The substitution of the conserved cysteine residues results in inability of covalently bound flavins to stabilize flavosemiquinone states, i.e. leads to incorrect folding of the NQR complex
C120G
NqrE-C120G. The substitution of the conserved cysteine residue in NqrE subunit of the enzyme blocks the Na+-dependent and HQNO-sensitive quinone reductase activity of the enzyme, being without effect on the interaction of the enzyme with reduced nicotinamide hypoxanthine dinucleotide and menadione. The substitution of the conserved cysteine residues results in inability of covalently bound flavins to stabilize flavosemiquinone states, i.e. leads to incorrect folding of the NQR complex
C26G
NqrE-C26G. The substitution of the conserved cysteine residue in NqrE subunits of the enzyme blocks the Na+-dependent and HQNO-sensitive quinone reductase activity of the enzyme, being without effect on the interaction of the enzyme with reduced nicotinamide hypoxanthine dinucleotide and menadione. The substitution of the conserved cysteine residues results in inability of covalently bound flavins to stabilize flavosemiquinone states, i.e. leads to incorrect folding of the NQR complex
C29A
NqrD-C29A. The substitutions of the conserved cysteine residue in NqrD subunit of the enzyme blocks the Na+-dependent and HQNO-sensitive quinone reductase activity of the enzyme, being without effect on the interaction of the enzyme with reduced nicotinamide hypoxanthine dinucleotide and menadione. The substitution of the conserved cysteine residues results in inability of covalently bound flavins to stabilize flavosemiquinone states, i.e. leads to incorrect folding of the NQR complex
C70A
C76A
C79A
D133A
The mutants NqrB-E28A, NqrB-E144L, NqrD-D133A, and NqrD-D88L, in which sodium stimulation of sodium-dependent quinone reductase activity activity is lowered significantly but not eliminated, produces diminished levels of sodium pumping activity
D346A
D397A
D397C
D397E
D397K
D397N
D397S
D88L
mutant enzyme shows little or no change in the KmappNa+ for sodium. The mutant enzyme in which sodium stimulation of sodium-dependent quinone reductase activity activity is lowered significantly but not eliminated, produces diminished levels of sodium pumping activity
E144C
mutation in subunit NrqB, the inhibitory effects of and 3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate are almost completely abolished, while their the binding reactivities are unchanged
E144L
NqrB-E144L. The mutant in which sodium stimulation of sodium-dependent quinone reductase activity is lowered significantly but not eliminated, produces diminished levels of sodium pumping activity
E28A
NqrB-D346A. The mutant in which sodium stimulation of sodium-dependent quinone reductase activity is lowered significantly but not eliminated, produces diminished levels of sodium pumping activity. Little or no change in the KmappNa+ for sodium
E95A
NqrE-E95A. The effect of sodium on sodium-dependent quinone reductase activity is almost eliminated. Sodium-dependent quinone reductase activity of mutant enzyme is very similar to that of wild-type Na+-NQR in the absence of sodium. The mutant enzyme in which the sodium-dependent quinone reductase activity activity is insensitive to sodium is unable to form a sodium gradient
G140A
G140L
G141A
G141L
G141V
G38V
mutation in subunit NrqA, no significant changes in the effects of inhibitors tested
H216L
the subunit NqrC mutant contains 0.2-0.3 mol of FMN per mol of protein, the mutation reduces the FMN content (30%) of the isolated subunit NqrC
R210L
FAD mutant of subunit NqrF with severely reduced activity using ubiquinone-1or ferricyanide and NADH as substrates
S246A
T225L
no covalent flavin is detected when threonine-225 is replaced by leucine, the mutation eliminates flavin binding by subunit NqrC
T225Y
T236Y
T236Y/T225Y
Y212L
Y36A
mutation in subunit NrqA, no significant changes in the effects of inhibitors tested
additional information
C111A
subunit NqrF, 3% of the NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
C111A
subunit NqrF, 2Fe-2S center mutant with severely reduced activity using ubiquinone-1 and NADH as substrates. NADH:ferricyanide oxidoreduction activity remains unchanged compared to the wild type enzyme
C70A
subunit NqrF, 3% of the NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
C70A
subunit NqrF, 2Fe-2S center mutant with severely reduced activity using ubiquinone-1 and NADH as substrates, the amount of FAD in the C70A mutant is essentially the same as in the wild type. NADH:ferricyanide oxidoreduction activity remains unchanged compared to the wild type enzyme
C76A
subunit NqrF, 3% of the NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
C76A
-
in the NqrF-C76A mutant, turnover with ferricyanide is almost the same as in the wild type enzyme
C76A
-
in this mutant one of the cysteine ligands of the 2Fe-2S center is changed to alanine, resulting in the complete loss of this cofactor. In all three flavin mutants (NqrB-T236Y, NqrCT225Y, and the double mutant) as well as the 2Fe-2S center mutant (NqrF-C76A), the reduction by NADH of the neutral flavosemiquinone is dramatically slowed, or abolished, compared with the wild type enzyme
C76A
-
mutant lacks the 2Fe-2S center in NqrF (NqrF-C76A). The reaction of the mutant enzyme with NADH consists of a single phase, whether or not Na+ is present. Turnover with ferricyanide is almost the same as in the wild type enzyme. Two electron reduction of a flavin is the only significant phase remaining from the wild type reaction, allows the redox carrier upstream of the 2Fe-2S center to be assigned to the first kinetic phase of the wild type reaction and hence to the FAD in NqrF. Reduction by NADH of the neutral flavosemiquinone is strongly slowed or abolished, compared with the wild type enzyme
C76A
subunit NqrF, 2Fe-2S center mutant with severely reduced activity using ubiquinone-1 and NADH as substrates. NADH:ferricyanide oxidoreduction activity remains unchanged compared to the wild type enzyme
C76A
-
mutant of subunit NqrF which lacks the 2Fe-2S-center cofactor and shows a 90% decrease in menadione reductase activity
C79A
subunit NqrF, 3% of the NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
C79A
subunit NqrF, 2Fe-2S center mutant with severely reduced activity using ubiquinone-1 and NADH as substrates. NADH:ferricyanide oxidoreduction activity remains unchanged compared to the wild type enzyme
D346A
NqrB-D397A. Sodium-dependent quinone reductase activity of the mutant enzyme is very similar to that of wild-type Na+-NQR in the absence of sodium. The mutant enzyme in which the sodium-dependent quinone reductase activity activity is insensitive to sodium is unable to form a sodium gradient.
D346A
the effect of sodium on sodium-dependent quinone reductase activity is almost eliminated. Little or no change in the KmappNa+ for sodium
D346A
-
the redox reaction in this mutant is significantly inhibited and the Na+ sensitivity of CoQ reduction is completely abolished, while the apparent affinity for Na+ is essentially unchanged. In D346A, electron transfer between FMN from subunit NqrB and riboflavin is inhibited
D397A
sodium-dependent quinone reductase activity of mutant enzyme is very similar to that of wild-type Na+-NQR in the absence of sodium. The mutant enzyme in which the sodium-dependent quinone reductase activity is insensitive to sodium is unable to form a sodium gradient
D397A
the effect of sodium on sodium-dependent quinone reductase activity is almost eliminated
D397A
-
the reduction of subunit NqrB mutant D397A by NADH is very similar to the reduction of wild type enzyme but does not show sensitivity to Na+, and the rate of the stable neutral riboflavin radical to riboflavin-H2 is decreased 3fold. In NqrB-D397A, the reaction takes place in two steps: rapid oxidation of riboflavin and the two FMNs, followed by oxidation of FAD, at a 16fold lower rate
D397A
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site, the kinetics of the NqrB-D397A mutant show an unsaturable behavior for sodium in contrast to the Michaelis-Menten kinetics of the wild-type enzyme. The mutant is not stimulated by lithium
D397A
subunit NqrB, mutant is completely insensitive to sodium and also not stimulated by lithium
D397C
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site
D397C
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site, the mutation that eliminates half of the negative charge, is stimulated only 2.6fold by sodium
D397C
subunit NqrB, mutant is stimulated by Li+. Mutation results in negative cooperativity
D397E
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site
D397E
subunit NqrB, mutant is stimulated by Li+. Km value is not affected. Mutant contains approximately the same conformational flexibility as the wild type enzyme and is able to undergo a series of conformational changes induced by the redox reaction and by the addition of different cations
D397E
subunit NqrB. In the mutant, the spectral features characteristic of COOH groups are shifted, and the hydrogen binding of the ion binding cluster is weakened
D397K
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site, the introduction of a positive charge abates completely the stimulatory effect of sodium, the mutant is not stimulated by lithium
D397K
subunit NqrB, mutant is completely insensitive to sodium and also not stimulated by lithium
D397N
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site, the mutation that eliminates the negative charge, but that introduces a polar residue with a partial negative charge, is stimulated only 2fold by sodium
D397N
subunit NqrB, mutant is stimulated by Li+. Km value is not affected. Sodium-binding site II is inactive
D397S
site-directed mutagenesis of subunit NqrB residue, part of the sodium binding site, the mutation that eliminates the negative charge, but that introduces a polar residue with a partial negative charge, is stimulated only 2fold by sodium
D397S
subunit NqrB, mutant is stimulated by Li+. Km value is not affected
G140A
subunit NqrB. Site-directed mutagenesis, the mutant shows highly reduced activity and reduced inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide compared to the wild-type enzyme
G140A
sodium-pumping activity of the mutant is not affected under partial turnover conditions. Mutation affects exclusively the binding of ubiquinone
G140A
mutation in subunit NrqB, affects ubiquinone reduction and the proper functioning of the sodium pump. Mutation does not not affect the dissociation constant of ubiquinone or inhibitor 2-n-heptyl-4-hydroxyquinoline N-oxide from Na+-NQR and prevents the conformational change involved in ubiquinone binding
G140A
mutation in subunit NrqB, the inhibitory effects of 5-azido-3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 4-iodobenzoate and 3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate are almost completely abolished, while their the binding reactivities are unchanged
G140L
subunit NqrB, site-directed mutagenesis. The mutant shows highly reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme
G140L
mutant shows nonsaturating behavior with up to 50 microM ubiquinone
G141A
subunit NqrB, site-directed mutagenesis. The mutant shows reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme
G141A
overall structure of the enzyme is not disturbed by the mutation. Mutation specifically affects the ubiquinone binding site. 6fold increase in Km value
G141A
mutation in subunit NrqB, decrease in the inhibitory effects of koromicin and 3-[(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl]-2-methylquinolin-4(1H)-one 2-azido-5-iodobenzoate
G141L
3fold increase in Km value
G141L
subunit NqrB, site-directed mutagenesis. The mutant shows reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme
G141V
site-directed mutagenesis, the mutant shows reduced activity and reduced inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide compared to the wild-type enzyme
G141V
9fold increase in Km value
G141V
mutation in subunit NrqB, affects ubiquinone reduction and the proper functioning of the sodium pump. Mutation does not not affect the dissociation constant of ubiquinone or inhibitor 2-n-heptyl-4-hydroxyquinoline N-oxide from Na+-NQR and prevents the conformational change involved in ubiquinone binding
S246A
subunit NqrF, extremely low NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
S246A
-
the wild type enzyme is capable of transferring electrons from NADH to ferricyanide. This reaction is almost completely abolished in the NqrFS246A mutant, which lacks the FAD
S246A
-
this reaction is almost completely abolished in the NqrFS246A mutant, which lacks the FAD
S246A
FAD mutant of subunit NqrF with severely reduced activity possessing only a small residual amount of FAD using ubiquinone-1 or ferricyanide and NADH as substrates
T225Y
-
lacks FMN in subunit NqrC, is not able to carry out electron transfer from NADH to ubiquinone, but instead from NADH to ferricyanide
T225Y
-
in all three flavin mutants (NqrB-T236Y, NqrCT225Y, and the double mutant) as well as the 2Fe-2S center mutant (NqrF-C76A), the reduction by NADH of the neutral flavosemiquinone is dramatically slowed, or abolished, compared with the wild type enzyme
T225Y
-
mutant lacks FMN in subunit C (NqrC-T225Y). Reduction by NADH of the neutral flavosemiquinone is strongly slowed or abolished, compared with the wild type enzyme
T225Y
-
the mutant lacks FMN from subunit NqrC and is unable to produce an electrochemical potential under either partial or multiple turnover conditions
T236Y
-
lacks FMN in subunit NqrB, is not able to carry out electron transfer from NADH to ubiquinone, but instead from NADH to ferricyanide
T236Y
-
in all three flavin mutants (NqrB-T236Y, NqrCT225Y, and the double mutant) as well as the 2Fe-2S center mutant (NqrF-C76A), the reduction by NADH of the neutral flavosemiquinone is dramatically slowed, or abolished, compared with the wild type enzyme
T236Y
-
mutant lacks FMN in subunit B (NqrB-T236Y). Reduction by NADH of the neutral flavosemiquinone is strongly slowed or abolished, compared with the wild type enzyme
T236Y
-
the mutant lacks FMN from subunit NqrB and is unable to produce an electrochemical potential under either partial or multiple turnover conditions
T236Y/T225Y
-
in all three flavin mutants (NqrB-T236Y, NqrCT225Y, and the double mutant) as well as the 2Fe-2S center mutant (NqrF-C76A), the reduction by NADH of the neutral flavosemiquinone is dramatically slowed, or abolished, compared with the wild type enzyme
T236Y/T225Y
-
mutant lacks FMN in subunit B and FMN in subunit C (NqrB-T236Y/NqrC-T225Y). This mutant contains only the noncovalently bound FAD in NqrF and the noncovalently bound riboflavin. The reaction of the mutant enzyme with NADH consists of a single phase, whether or not Na+ is present. The rate constants are almost identical, are as the spectra. The spectra appear to show two-electron reduction of a flavin. The spectra and rates are consistent with those of the first phase of the reaction of the wild type enzyme in the absence of Na+. Reduction by NADH of the neutral flavosemiquinone is strongly slowed or abolished, compared with the wild type enzyme
T236Y/T225Y
-
NqrB-T236Y/NqrC-T225Y, double mutant that lacks both covalently bound FMN cofactors. The double mutant contains riboflavin and FAD in a 0.6:1 ratio, as the only flavins in the enzyme, noncovalently bound flavins are detected. In the oxidized form, the double mutant exhibits an EPR signal consistent with a neutral flavosemiquinone radical, which is abolished on reduction of the enzyme
Y212L
subunit NqrF, extremely low NADH:ubiquinone-1 oxidoreductase activity compared to wild-type enzyme
Y212L
FAD mutant of subunit NqrF with severely reduced activity using ubiquinone-1 or ferricyanide and NADH as substrates
additional information
-
a soluble variant of NqrF lacking its hydrophobic, N-terminal helix (NqrF') is produced in Vibrio cholerae wild type and nqr deletion strain. Under identical conditions of growth and induction, the yield of NqrF' increased by 30% in the presence of the Na+-NQR. FAD-containing NqrF' species with or without the FeS cluster are observed, indicating that assembly of the FeS center, but not insertion of the flavin cofactor, is limited during overproduction in Vibrio cholerae. NqrF' lacking the [2Fe2S] cluster is less stable, partially unfolded, and therefore prone to proteolytic degradation in Vibrio cholerae
additional information
construction of a Vibrio cholerae deletion strain lacking the genomic nqr operon, DELTAnqr
additional information
preparation of membranes containing subcomplexes of Na+-NQR, NqrBCDEF, NqrACDEF, NqrABDEF, NqrABCEF, and NqrABCF, by expressing mutant forms of the Na+-NQR in Vibrio cholerae enzyme-deficient mutant strain DELTAnqr
additional information
residue D397 in subunit NqrB forms part of one of the at least two sodium-binding sites. Both size and charge at this position are critical for the function of the enzyme. The residue is involved in cation selectivity, has a critical role in the communication between sodium-binding sites, by promoting cooperativity, and controls the electron transfer step involved in sodium uptake
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
His6-tagged wild-type and mutant forms of the Na+-NQR from Vibrio cholerae mutant strain DELTAnqr by nickel affinity chromatography and gel filtration
mutant enzyme E95A
mutant enzymes NqrB-D346A and NqrB-D397A
Ni-NTA agarose column chromatography and Superdex 200 gel filtration
-
Ni-NTA column chromatography
Ni-NTA column chromatography and Mono Q HR 10/10 column chromatography
-
Ni-NTA column chromatography and Sephadex G-25 gel filtration
recombinant enzyme
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography
recombinant N-terminally His6-tagged full-length enzyme and large subunit NqrA from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
recombinant N-terminally His6-tagged monomeric large subunit NqrA from Vibrio cholerae strain O395 N1 DELTAnqr membranes by ultracentrifugation, nickel affinity chromatography, and gel filtration in the presence of 0.05% w/v n-dodecyl-beta-D-maltoside
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloned under the regulation of the P(BAD) promoter, successfully expressed in Vibrio cholerae
-
cloned under the regulation of the PBAD promoter and expressed in Vibrio cholera
-
expressed in Vibrio cholerae strain O395N1-toxT:lac::DELTAnqrA-F
expression in Escherichia coli
expression of full-length enzyme and N-terminally His6-tagged large subunit NqrA in Vibrio cholerae strain O395 N1 DELTAnqr
-
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression of N-terminally His6-tagged full-length enzyme and large subunit NqrA in Escherichia coli strain BL21(DE3)
-
hexahistidine-tagged Na+-NQR is expressed in Vibrio cholerae strain DELTAnqr lacking expression of the NQR complex
-
subunit NqrC is expressed in Vibrio cholerae and in Escherichia coli TOP10 cells. Subunit NqrB expression is toxic for Vibrio cholerae
wild-type and mutant forms of the Na+-NQR from Vibrio cholerae as recombinant proteins containing a His6-tag at the N-terminus of subunit NqrA in an enzyme-deficient mutant strain DELTAnqr of Vibrio cholerae
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
reconstitution of Na+-NQR in proteoliposomes and generation of membrane potential. Purified Na+-NQR is mixed with Escherichia coli phospholipids and n-octyl glucoside (detergent/phospholipid ratio = 1.3) in buffer containing 100 mM KCl, 50 mM HEPES, 1 mM EDTA, pH 7.0. The detergent is removed slowly
-
reconstitution of purified recombinant enzyme with riboflavin, enzyme is mixed with 0.1 mM riboflavin from a1 mM riboflavin stock solution in N,N-dimethylformamide, 10 min, 25°C
reconstitution of recombinant Na+-NQR and recombinant His6-tagged NqrA with ubiquinone-8, overview
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
-
Vibrio cholerae Na+-NQR is significant for the induction of virulence factors. Thus, this enzyme can be used as a target in the treatment or prevention of many infectious diseases
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Barquera, B.; Hellwig, P.; Zhou, W.; Morgan, J.E.; Hse, C.C.; Gosink, K.K.; Nilges, M.; Bruesehoff, P.J.; Roth, A.; Lancaster, C.R.D.; Gennis, R.B.
Purification and characterization of the recombinant Na+-translocating NADH:quinone oxidoreductase from Vibrio cholera
Biochemistry
41
3781-3789
2002
Vibrio cholerae serotype O1
Barquera, B.; Nilges, M.J.; Morgan, J.E.; Ramirez-Silva, L.; Zhou, W.; Gennis, R.B.
Mutagenesis study of the 2Fe-2S center and the FAD binding site of the Na(+)-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae
Biochemistry
43
12322-12330
2004
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Barquera, B.; Morgan, J.E.; Lukoyanov, D.; Scholes, C.P.; Gennis, R.B.; Nilges, M.J.
X- and W-band EPR and Q-band ENDOR studies of the flavin radical in the Na+ -translocating NADH:quinone oxidoreductase from Vibrio cholerae
J. Am. Chem. Soc.
125
265-275
2003
Vibrio cholerae serotype O1
Trk, K.; Puhar, A.; Neese, F.; Bill, E.; Fritz, G.; Steuber, J.
NADH oxidation by the Na+-translocating NADH:quinone oxidoreductase from Vibrio cholerae: functional role of the NqrF subunit
J. Biol. Chem.
279
21349-21355
2004
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Tao, M.; Tuerk, K.; Diez, J.; Gruetter, M.G.; Fritz, G.; Steuber, J.
Crystallization of the NADH-oxidizing domain of the Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae
Acta Crystallogr. Sect. F
F62
110-112
2006
Vibrio cholerae serotype O1
Lin, P.C.; Puhar, A.; Tuerk, K.; Piligkos, S.; Bill, E.; Neese, F.; Steuber, J.
A vertebrate-type ferredoxin domain in the Na+-translocating NADH dehydrogenase from Vibrio cholerae
J. Biol. Chem.
280
22560-22563
2005
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Barquera, B.; Ramirez-Silva, L.; Morgan, J.E.; Nilges, M.J.
A new flavin radical signal in the Na(+)-pumping NADH:quinone oxidoreductase from Vibrio cholerae. An EPR/electron nuclear double resonance investigation of the role of the covalently bound flavins in subunits B and C
J. Biol. Chem.
281
36482-36491
2006
Vibrio cholerae serotype O1
Fadeeva, M.S.; Bertsova, Y.V.; Verkhovsky, M.I.; Bogachev, A.V.
Site-directed mutagenesis of conserved cysteine residues in NqrD and NqrE subunits of Na+-translocating NADH:quinone oxidoreductase
Biochemistry (Moscow)
73
123-129
2008
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Juarez, O.; Athearn, K.; Gillespie, P.; Barquera, B.
Acid residues in the transmembrane helices of the Na+-pumping NADH:quinone oxidoreductase from vibrio cholerae involved in sodium translocation
Biochemistry
48
9516-9524
2009
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Tao, M.; Casutt, M.S.; Fritz, G.; Steuber, J.
Oxidant-induced formation of a neutral flavosemiquinone in the Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae
Biochim. Biophys. Acta
1777
696-702
2008
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Juarez, O.; Nilges, M.J.; Gillespie, P.; Cotton, J.; Barquera, B.
Riboflavin is an active redox cofactor in the Na+-pumping NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae
J. Biol. Chem.
283
33162-33167
2008
Vibrio cholerae serotype O1
Juarez, O.; Morgan, J.E.; Barquera, B.
The electron transfer pathway of the Na+-pumping NADH:Quinone Oxidoreductase from Vibrio cholerae
J. Biol. Chem.
284
8963-8972
2009
Vibrio cholerae serotype O1
Tao, M.; Fritz, G.; Steuber, J.
The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae enhances insertion of FeS in overproduced NqrF subunit
J. Inorg. Biochem.
102
1366-1372
2008
Vibrio cholerae serotype O1
Casutt, M.S.; Wendelspiess, S.; Steuber, J.; Fritz, G.
Crystallization of the Na+-translocating NADH:quinone oxidoreductase from Vibrio cholerae
Acta Crystallogr. Sect. F
66
1677-1679
2010
Vibrio cholerae serotype O1
Bogachev, A.V.; Verkhovsky, M.I.
Na+-translocating NADH:quinone oxidoreductase: progress achieved and prospects of investigations
Biochemistry
70
143-149
2005
Haemophilus influenzae, Klebsiella pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis, Porphyromonas gingivalis, Pseudomonas aeruginosa, Shewanella putrefaciens, Vibrio alginolyticus, Vibrio cholerae serotype O1, Vibrio harveyi, Yersinia pestis
Verkhovsky, M.I.; Bogachev, A.V.
Sodium-translocating NADH:quinone oxidoreductase as a redox-driven ion pump
Biochim. Biophys. Acta
1797
738-746
2010
Azotobacter vinelandii, Haemophilus influenzae, Klebsiella pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis, Porphyromonas gingivalis, Pseudomonas aeruginosa, Vibrio alginolyticus, Vibrio cholerae serotype O1, Vibrio harveyi, Yersinia pestis
Barquera, B.; Hase, C.C.; Gennis, R.B.
Expression and mutagenesis of the NqrC subunit of the NQR respiratory Na+ pump from Vibrio cholerae with covalently attached FMN
FEBS Lett.
492
45-49
2001
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Lin, P.C.; Turk, K.; Hase, C.C.; Fritz, G.; Steuber, J.
Quinone reduction by the Na+-translocating NADH dehydrogenase promotes extracellular superoxide production in Vibrio cholerae
J. Bacteriol.
189
3902-3908
2007
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Juarez, O.; Morgan, J.E.; Nilges, M.J.; Barquera, B.
Energy transducing redox steps of the Na+-pumping NADH:quinone oxidoreductase from Vibrio cholerae
Proc. Natl. Acad. Sci. USA
107
12505-12510
2010
Vibrio cholerae serotype O1
Casutt, M.S.; Huber, T.; Brunisholz, R.; Tao, M.; Fritz, G.; Steuber, J.
Localization and function of the membrane-bound riboflavin in the Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae
J. Biol. Chem.
285
27088-27099
2010
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Juarez, O.; Shea, M.E.; Makhatadze, G.I.; Barquera, B.
The role and specificity of the catalytic and regulatory cation-binding sites of the Na+-pumping NADH:quinone oxidoreductase from Vibrio cholerae
J. Biol. Chem.
286
26383-26390
2011
Vibrio cholerae serotype O1
Casutt, M.S.; Nedielkov, R.; Wendelspiess, S.; Vossler, S.; Gerken, U.; Murai, M.; Miyoshi, H.; Moeller, H.M.; Steuber, J.
Localization of ubiquinone-8 in the Na+-pumping NADH:quinone oxidoreductase from Vibrio cholerae
J. Biol. Chem.
286
40075-40082
2011
Vibrio cholerae serotype O1
Juarez, O.; Neehaul, Y.; Turk, E.; Chahboun, N.; DeMicco, J.M.; Hellwig, P.; Barquera, B.
The role of glycine residues 140 and 141 of subunit B in the functional ubiquinone binding site of the Na+-pumping NADH:quinone oxidoreductase from Vibrio cholerae
J. Biol. Chem.
287
25678-25685
2012
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Nedielkov, R.; Steffen, W.; Steuber, J.; Moller, H.M.
NMR reveals double occupancy of quinone-type ligands in the catalytic quinone binding site of the Na+-translocating NADH:quinone oxidoreductase from Vibrio cholerae
J. Biol. Chem.
288
30597-30606
2013
Vibrio cholerae serotype O1
Shea, M.E.; Juarez, O.; Cho, J.; Barquera, B.
Aspartic acid 397 in subunit B of the Na+-pumping NADH: quinone oxidoreductase from Vibrio cholerae forms part of a sodium binding site, is involved in cation selectivity and affects cation binding site cooperativity
J. Biol. Chem.
288
31241-31249
2013
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Casutt, M.S.; Schlosser, A.; Buckel, W.; Steuber, J.
The single NqrB and NqrC subunits in the Na(+)-translocating NADH: quinone oxidoreductase (Na(+)-NQR) from Vibrio cholerae each carry one covalently attached FMN
Biochim. Biophys. Acta
1817
1817-1822
2012
Vibrio cholerae serotype O1 (A5F5X0), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X0)
Minato, Y.; Fassio, S.R.; Kirkwood, J.S.; Halang, P.; Quinn, M.J.; Faulkner, W.J.; Aagesen, A.M.; Steuber, J.; Stevens, J.F.; Haese, C.C.
Roles of the sodium-translocating NADH:quinone oxidoreductase (Na+-NQR) on Vibrio cholerae metabolism, motility and osmotic stress resistance
PLoS ONE
9
e97083
2014
Vibrio cholerae serotype O1
Vohl, G.; Nedielkov, R.; Claussen, B.; Casutt, M.; Vorburger, T.; Diederichs, K.; Muller, H.; Steuber, J.; Fritz, G.
Crystallization and preliminary analysis of the NqrA and NqrC subunits of the Na+-translocating NADH ubiquinone oxidoreductase from Vibrio cholerae
Acta Crystallogr. Sect. F
70
987-992
2014
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
-
Neehaul, Y.; Juarez, O.; Barquera, B.; Hellwig, P.
Infrared spectroscopic evidence of a redox-dependent conformational change involving ion binding residue NqrB-D397 in the Na+-pumping NADH quinone oxidoreductase from Vibrio cholerae
Biochemistry
52
3085-3093
2013
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
-
Vorburger, T.; Nedielkov, R.; Brosig, A.; Bok, E.; Schunke, E.; Steffen, W.; Mayer, S.; Gtz, F.; Mller, H.; Steuber, J.
Role of the Na+-translocating NADH quinone oxidoreductase in voltage generation and Na+ extrusion in Vibrio cholerae
Biochim. Biophys. Acta
1857
473-482
2016
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Steuber, J.; Vohl, G.; Muras, V.; Toulouse, C.; Claussen, B.; Vorburger, T.; Fritz, G.
The structure of Na+-translocating of NADH ubiquinone oxidoreductase of Vibrio cholerae Implications on coupling between electron transfer and Na+ transport
Biol. Chem.
396
1015-1030
2015
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Toulouse, C.; Claussen, B.; Muras, V.; Fritz, G.; Steuber, J.
Strong pH dependence of coupling efficiency of the Na+ - translocating NADH quinone oxidoreductase (Na+-NQR) of Vibrio cholerae
Biol. Chem.
398
251-260
2017
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Muras, V.; Dogaru-Kinn, P.; Minato, Y.; Haese, C.C.; Steuber, J.
The Na+-translocating NADH quinone oxidoreductase enhances oxidative stress in the cytoplasm of Vibrio cholerae
J. Bacteriol.
198
2307-2317
2016
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
Strickland, M.; Juarez, O.; Neehaul, Y.; Cook, D.A.; Barquera, B.; Hellwig, P.
The conformational changes induced by ubiquinone binding in the Na+-pumping NADH ubiquinone oxidoreductase (Na+-NQR) are kinetically controlled by conserved glycines 140 and 141 of the NqrB subunit
J. Biol. Chem.
289
23723-23733
2014
Vibrio cholerae serotype O1 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4), Vibrio cholerae serotype O1 ATCC 39541 (A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4)
Ito, T.; Murai, M.; Ninokura, S.; Kitazumi, Y.; Mezic, K.; Cress, B.; Koffas, M.; Morgan, J.; Barquera, B.; Miyoshi, H.
Identification of the binding sites for ubiquinone and inhibitors in the Na+-pumping NADH-ubiquinone oxidoreductase from Vibrio cholerae by photoaffinity labeling
J. Biol. Chem.
292
7727-7742
2017
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
-
Steuber, J.; Vohl, G.; Casutt, M.S.; Vorburger, T.; Diederichs, K.; Fritz, G.
Structure of the V. cholerae Na+-pumping NADH quinone oxidoreductase
Nature
516
62-67
2014
Vibrio cholerae serotype O1 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8), Vibrio cholerae serotype O1 ATCC 39315 (Q9KPS1 and Q9KPS2 and P0C6E0 and Q9X4Q6 and Q9X4Q7 and Q9X4Q8)
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Transporter Classification Database (TCDB): 3.D.5.1.1
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