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Information on EC 7.4.2.5 - bacterial ABC-type protein transporter and Organism(s) Escherichia coli and UniProt Accession P10408
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7.4.2.5 bacterial ABC-type protein transporterIUBMB Comments
An ATP-binding cassette (ABC) type transporter, characterized by the presence of two similar ATP-binding domains/proteins and two integral membrane domains/proteins. This entry stands for a family of bacterial enzymes that are dedicated to the secretion of one or several closely related proteins belonging to the toxin, protease and lipase families. Examples from Gram-negative bacteria include alpha-hemolysin, cyclolysin, colicin V and siderophores, while examples from Gram-positive bacteria include bacteriocin, subtilin, competence factor and pediocin.
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Word Map
- 7.4.2.5
-
caco-2
- tripeptides
- jejunum
-
gly-sar
-
glycylsarcosine
-
secyeg
- xenopus
- oocyte
-
histocompatibility
-
multidrug
-
prodrugs
- p-glycoprotein
-
enterocytes
- duodenum
-
peptidomimetic
- beta-lactams
- preproteins
-
basolateral
- ileum
- cephalexin
- laevis
-
peptide-like
-
translocons
-
transepithelial
-
brush-border
-
carrier-mediated
-
h+-coupled
-
villus
-
electrogenic
-
pepts
-
proton-dependent
- valacyclovir
- acyclovir
-
transporter-mediated
-
tapasin
- carnosine
-
sec-dependent
-
paracellular
-
membrane-embedded
-
two-electrode
- bestatin
- oct1
-
oatp2b1
-
oatps
-
antigen-processing
- gly-gly
- cefixime
-
i-restricted
-
apical-to-basolateral
-
serine-rich
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
pept1, peptide transporter, pept2, seca2, abc transport, seca protein, peptide transporter 1, abcb10, seca1, seca atpase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
peptide-transporting ATPase
-
-
-
-
SecA
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric ester
transmembrane transport
transmembrane transport
hydolysis of phosphoric ester
-
-
-
-
transmembrane transport
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (ABC-type, peptide-exporting)
An ATP-binding cassette (ABC) type transporter, characterized by the presence of two similar ATP-binding domains/proteins and two integral membrane domains/proteins. This entry stands for a family of bacterial enzymes that are dedicated to the secretion of one or several closely related proteins belonging to the toxin, protease and lipase families. Examples from Gram-negative bacteria include alpha-hemolysin, cyclolysin, colicin V and siderophores, while examples from Gram-positive bacteria include bacteriocin, subtilin, competence factor and pediocin.
CAS REGISTRY NUMBER
COMMENTARY
9000-83-3
-
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
ATP + H2O + alpha-haemolysin/in
ADP + phosphate + alpha-haemolysin/out
-
-
-
-
?
ATP + H2O + beta-L-Ala-L-Lys-7-amido-4-methylcoumarin-3-acetic acid/in
ADP + phosphate + beta-L-Ala-L-Lys-7-amido-4-methylcoumarin-3-acetic acid/out
-
-
-
?
additional information
?
-
-
one intact nucleotide-binding domain within a dimer is sufficient for ATP hydrolysis. One ATP-binding site of the dimer is able to function independently of the hydrolytic capability of the neighboring ATP-binding site. Sequential mechanism of ATP hydrolysis in the intact HlyB transporter
-
-
?
ATP + H2O
ADP + phosphate
-
ATP hydrolysis by SecA is required only if periplasmic loops larger than 30 amino acids have to be translocated. The signal recognition particle-dependent and SecA-dependent multiple spanning membrane protein YidC becomes SecA-independent if the large periplasmic loop connecting transmembrane domains 1 and 2 is reduced to less than 30 amino acids. The SecA dependence of a bacterial membrane protein is not solely determined by the length of the periplasmic loop but also by the presence of a down-stream TM domain
-
-
?
ATP + H2O
ADP + phosphate
-
ATPase SecA provides the driving force for the transport of secretory proteins across the cytoplasmic membrane of Escherichia coli
-
-
?
ATP + H2O
ADP + phosphate
-
SEcA drives a constant rate of preprotein translocation consistent with a stepping mechanism of translocation. The delay in full-length translocation for longer preproteins is dependent on the SecA motor function. The rate of protein translocation is controlled by the ATP concentration and is independent from the length of the preprotein
-
-
?
ATP + H2O
ADP + phosphate
-
SecB, a small cytosolic chaperone, captures the precursor polypeptides before they fold and delivers them to the membrane translocon through interactions with SecA. Both SecB and SecA display twofold symmetry and yet the complex between the two is stabilized by contacts that are distributed asymmetrically
-
-
?
ATP + H2O
ADP + phosphate
-
the preprotein binding domain is a preprotein receptor and a physical bridge connecting bound preproteins to the DEAD motor
-
-
?
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
ATP + H2O + alpha-haemolysin/in
ADP + phosphate + alpha-haemolysin/out
-
-
-
-
?
ATP + H2O
ADP + phosphate
-
ATP hydrolysis by SecA is required only if periplasmic loops larger than 30 amino acids have to be translocated. The signal recognition particle-dependent and SecA-dependent multiple spanning membrane protein YidC becomes SecA-independent if the large periplasmic loop connecting transmembrane domains 1 and 2 is reduced to less than 30 amino acids. The SecA dependence of a bacterial membrane protein is not solely determined by the length of the periplasmic loop but also by the presence of a down-stream TM domain
-
-
?
ATP + H2O
ADP + phosphate
-
ATPase SecA provides the driving force for the transport of secretory proteins across the cytoplasmic membrane of Escherichia coli
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-((4-azidobenzyl)thio)-4-(4-(benzyloxy)phenyl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile
good antimicrobial inhibition with MIC of 12.5 microM
2-((4-azidobenzyl)thio)-6-oxo-4-(4-phenoxyphenyl)-1,6-dihydropyrimidine-5-carbonitrile
good antimicrobial inhibition with MIC of 18.2 microM
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(4-bromophenyl)-5-cyano-4-oxopyrimidine)
-
-
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(biphenyl-4-yl)-5-cyano-4-oxopyrimidine)
-
compound does not show antimicrobial activity
2-(benzylsulfanyl)-4-(biphenyl-4-yl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile
-
exhibits most potent inhibition effects against strain NR698 with increased outer membrane permeability
erythrosin B
-
the potency in inhibiting the truncated SecA ATPase correlates with the ability to inhibit the biologically relevant protein translocation activity of SecA and also translates into antibacterial effects
Mg2+
-
2 mM Mg2+ blocks SecA ATPase activity almost completely, the inhibition is abolishable by binding of SecA to the protein channel SecYEG in proteoliposomes (1 micromol)
Rose bengal
-
the potency in inhibiting the truncated SecA ATPase correlates with the ability to inhibit the biologically relevant protein translocation activity of SecA and also translates into antibacterial effects
additional information
-
Insertion of charged amino acid residues into the preprotein proOmpA strongly inhibits SecA translocation ATPase activity. Stretches of positively charged residues are much stronger translocation inhibitors and suppressors of the preprotein-stimulated SecA ATPase activity than negatively charged residues.
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
DTT
effect on mutant N95CC, a large stimulation of ATPase activity is only observed when both proOmpA and DTT are present
proOmpA
effect on mutant N95CC, a large stimulation of ATPase activity is only observed when both proOmpA and DTT are present
-
Calcium
-
27-82 micromol Ca2+ enhances ATPase activity in presence of Escherichia coli PE/anionic phospholipid membranes up to 3-fold
SecM
-
SecA synthesized without the signal sequence on SecM is much less active on average than that synthesized in combination with normal SecM. The elongation arrest per se is not sufficient in terms of the SecA-activating ability of SecM. Targeting of nascent SecM to the membrane translocon is another crucial feature for SecM to enhance SEcA activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
ATP hydrolysis rate SecA wild-type 6.1 nmol phosphate/min/nmol enzyme
-
additional information
additional information
-
ATP hydrolysis rate SecA wild-type 6.1 nmol phosphate/min/nmol enzyme
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.05
alafosfalin
mutant K274I, pH 7.5, temperature not specified in the publication
0.07
alafosfalin
wild-type, pH 7.5, temperature not specified in the publication
0.25
alafosfalin
mutant M154K, pH 7.5, temperature not specified in the publication
0.28
alafosfalin
mutant V252E, pH 7.5, temperature not specified in the publication
0.38
alafosfalin
mutant L190V, pH 7.5, temperature not specified in the publication
0.45
alafosfalin
mutant F197I, pH 7.5, temperature not specified in the publication
0.96
alafosfalin
mutant L324V, pH 7.5, temperature not specified in the publication
0.08
L-Ala-L-Ala
mutant M154K, pH 7.5, temperature not specified in the publication
0.2
L-Ala-L-Ala
mutant K274I, pH 7.5, temperature not specified in the publication
0.2
L-Ala-L-Ala
mutant L190V, pH 7.5, temperature not specified in the publication
0.21
L-Ala-L-Ala
mutant V252E, pH 7.5, temperature not specified in the publication
0.21
L-Ala-L-Ala
wild-type, pH 7.5, temperature not specified in the publication
0.31
L-Ala-L-Ala
mutant F197I, pH 7.5, temperature not specified in the publication
0.36
L-Ala-L-Ala
mutant L324V, pH 7.5, temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0024
2-((4-azidobenzyl)thio)-4-(4-(benzyloxy)phenyl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile
Escherichia coli
37°C, pH not specified in the publication
0.0022
2-((4-azidobenzyl)thio)-6-oxo-4-(4-phenoxyphenyl)-1,6-dihydropyrimidine-5-carbonitrile
Escherichia coli
37°C, pH not specified in the publication
0.002 - 0.02
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(4-bromophenyl)-5-cyano-4-oxopyrimidine)
0.002 - 0.05
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(biphenyl-4-yl)-5-cyano-4-oxopyrimidine)
0.06
2-(benzylsulfanyl)-4-(biphenyl-4-yl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile
Escherichia coli
-
recombinant SecA, pH 7.6, 40°C
0.025
eosin Y
Escherichia coli
-
truncated SecA without the C-terminal regulatory domain, 40°C, pH 7.6
0.002
erythrosin B
Escherichia coli
-
truncated SecA without the C-terminal regulatory domain, 40°C, pH 7.6
0.0005
Rose bengal
Escherichia coli
-
truncated SecA without the C-terminal regulatory domain, pH 7.6, 40°C
0.002
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(4-bromophenyl)-5-cyano-4-oxopyrimidine)
Escherichia coli
-
EcN68, the N-terminal fragment of SecA without the C-terminal regulatory domain, pH 7.6, 40°C
0.02
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(4-bromophenyl)-5-cyano-4-oxopyrimidine)
Escherichia coli
-
recombinant SecA, pH 7.6, 40°C
0.002
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(biphenyl-4-yl)-5-cyano-4-oxopyrimidine)
Escherichia coli
-
EcN68, the N-terminal fragment of SecA without the C-terminal regulatory domain, pH 7.6, 40°C
0.05
2,2'-(alpha,alpha'-xylene)bis(sulfanediyl)bis-(6-(biphenyl-4-yl)-5-cyano-4-oxopyrimidine)
Escherichia coli
-
recombinant SecA, pH 7.6, 40°C
0.08
alafosfalin
Escherichia coli
mutant K274I, pH 7.5, temperature not specified in the publication
0.11
alafosfalin
Escherichia coli
wild-type, pH 7.5, temperature not specified in the publication
0.39
alafosfalin
Escherichia coli
mutant M154K, pH 7.5, temperature not specified in the publication
0.44
alafosfalin
Escherichia coli
mutant V252E, pH 7.5, temperature not specified in the publication
0.59
alafosfalin
Escherichia coli
mutant L190V, pH 7.5, temperature not specified in the publication
0.71
alafosfalin
Escherichia coli
mutant F197I, pH 7.5, temperature not specified in the publication
1.5
alafosfalin
Escherichia coli
mutant L324V, pH 7.5, temperature not specified in the publication
0.12
L-Ala-L-Ala
Escherichia coli
mutant M154K, pH 7.5, temperature not specified in the publication
0.31
L-Ala-L-Ala
Escherichia coli
mutant K274I, pH 7.5, temperature not specified in the publication
0.32
L-Ala-L-Ala
Escherichia coli
mutant L190V, pH 7.5, temperature not specified in the publication
0.33
L-Ala-L-Ala
Escherichia coli
mutant V252E, pH 7.5, temperature not specified in the publication
0.33
L-Ala-L-Ala
Escherichia coli
wild-type, pH 7.5, temperature not specified in the publication
0.48
L-Ala-L-Ala
Escherichia coli
mutant F197I, pH 7.5, temperature not specified in the publication
0.56
L-Ala-L-Ala
Escherichia coli
mutant L324V, pH 7.5, temperature not specified in the publication
additional information
beta-Ala-Lys
Escherichia coli
IC50 value range between 0.1 and 1 mM for wild-type and E20-mutants, and increase to above 10 mM for mutant E388Q, pH not specified in the publication, temperature not specified in the publication
additional information
beta-Ala-Lys
Escherichia coli
-
IC50 value range between 0.1 and 1 mM for wild-type and E20-mutants, and increase to above 10 mM for mutant E388Q, pH not specified in the publication, temperature not specified in the publication
additional information
Gly-Lys
Escherichia coli
IC50 value range between 0.1 and 1 mM for wild-type and E20-mutants, and increase to about 2 mM for mutant E388Q, pH not specified in the publication, temperature not specified in the publication
additional information
Gly-Lys
Escherichia coli
-
IC50 value range between 0.1 and 1 mM for wild-type and E20-mutants, and increase to about 2 mM for mutant E388Q, pH not specified in the publication, temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
10 nM Pi/min/nM SecA in presence of Escherichia coli PE/anionic phospholipid membranes
additional information
-
425 mol ATP/min/mol SecA in presence of 0.2 micromol preprotein translocation substrate proOmpA in urea
additional information
-
in ATPase translocation activity assay with preprotein translocation substrate proOmpA: circa 0.8 mM ATP/2 micromol SecA after 1 min, mutant SecA proteins with deletions of amino acids between 2-11 shows ATPase translocation activity around 0.25 mM ATP/2 micromol SecA after 1 min
additional information
-
translocation ATPase activity: 0.9 nmol phosphate/microgramm SecA/min, mutants lacking 2 to 11 residues of the amino terminus of SecA shows defect translocation ATPase activities (below 0.1 nmol phosphate/microgramm/min)
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 8.5
-
pH 7.0: about 75% of maximal activity, pH 8.5: about 85% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
physiological function
-
Both SecA and the ribosome need to interact with the translocon during membrane protein insertion. SecA and ribosome binding to the translocon is mutually exclusive, implying that during membrane protein insertion, both ligands bind the translocon in a sequential manner
physiological function
-
multiple SecA molecules drive translocation across a single translocon with SecG inversion. Model of proOmpA translocation suggests that a single catalytic cycle of SecA causes translocation of 1013 kDa with ATP binding and hydrolysis, and SecG inversion is required when the next SecA cycle begins with additional ATP hydrolysis
physiological function
-
SecA ATPase associates with the SecY complex to push preproteins across the bacterial membrane. One SecY copy is sufficient to bind SecA and the preprotein, but only the SecY dimer together with acidic lipids supports the activation of the SecA translocation ATPase. In nanodiscs, the dimer is predominantly arranged in a back-to-back manner and remains active even if a constituent SecY copy is defective for SecA binding. In membrane vesicles and in intact cells, the coproduction of two inactive SecYs, one for channel gating and the other for SecA binding, recreates a functional translocation unit
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
97000
102000
2 * 102000, gel filtration, furthermore electron paramagnetic resonance spectroscopy used to identify the interactive binding surface of SecA
200000
-
molecular weight depends on dimerization rate which depends on concentration of SecA and KCl: at 100 nM KCl molecular weight is 200000 Da, at 300 nM KCl molecular weight is around 140000 Da (both at 1microM SecA), monomer-dimer equilibrium is altered in SecA mutants: mutants lacking 2 to 11 residues of the amino terminus of SecA failed to form dimers at 1microM SecA and 300 nM KCl, determination by gel filtration controlling eluate by photodiode array UV/Vis detector, a differential refractometer and a static, multiangle laser light scattering detector
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
monomer
additional information
dimer
-
is the active form of the enzyme. ATP-promoted dimerization of HlyB-nucleotide binding domain might involve formation of two interconvertible forms: a readily dissociable active dimer and a more stable, reversibly inactive form of the dimer
dimer
-
membrane-bound SEcA dimer exists and is required for effective protein translocation
dimer
2 * 102000, gel filtration, furthermore electron paramagnetic resonance spectroscopy used to identify the interactive binding surface of SecA
dimer
-
chemical cross-linking experiments, dimerization is necessary for the translocation ATPase activity of SecA, monomer-dimer equilibrium is altered in SecA mutants: mutants lacking 2 to 11 residues of the amino terminus of SecA failed to form dimers at 1microM SecA and 300 nM KCl, determination by gel filtration controlling eluate by photodiode array UV/Vis detector, a differential refractometer and a static, multiangle laser light scattering detector
monomer
-
under most conditions, only the monomer form can be detected, indicating that dimers are very unstable. Monomers can interact at least transiently and are the important species during ATP hydrolysis
additional information
the dimer, crosslinked by disulfide bridges, is inactive, the monomer is active
additional information
residues Leu6, the amino terminus (residues 2 to 11) in the nucleotide binding domain, Phe263 in the preprotein binding domain, and Tyr794 and Arg805 in the intramolecular regulator of the ATPase 1 domain are involved in ecSecA dimerization
additional information
-
components involved are two helper proteins and the ABC protein: membrane fusion protein (MFP), outer membrane protein (OMP), and the ABC protein
additional information
-
components involved are two helper proteins and the ABC protein: membrane fusion protein (MFP), outer membrane protein (OMP), and the ABC protein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
molecular docking of inhibitor 2-((4-azidobenzyl)thio)-4-(4-(benzyloxy)phenyl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile. The azido group points towards hydrophilic residues including A/Asp512, A/Arg509, A/Gly510 and A/Thr511. The torsion around the -CH2O- atoms between the two phenyl groups allows the terminal phenyl group to rest in a pocket away from the hydrophilic residues B/Thr511, B/Gly510 and B/Glu487. Most of the active inhibitors seem to bind at the interface of chains A and B
homology modeling based on the crystal structure of the Shewanella oneidensis peptide transporter PepTso, identifies Glu56 and Arg305 as potential periplasmic gating residues
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Cys-less
mutant for analyzing the interaction and regulatory domains of SecA
E400C
mutant for analyzing the interaction and regulatory domains of SecA
E400C/R642C
mutant for analyzing the interaction and regulatory domains of SecA
E400R
mutant for analyzing the interaction and regulatory domains of SecA
E400R/A628T
mutant for analyzing the interaction and regulatory domains of SecA
E400R/E619K
mutant for analyzing the interaction and regulatory domains of SecA
E400R/H620P
mutant for analyzing the interaction and regulatory domains of SecA
E400R/I627T
mutant for analyzing the interaction and regulatory domains of SecA
E400R/L610P
mutant for analyzing the interaction and regulatory domains of SecA
E400R/M607T
mutant for analyzing the interaction and regulatory domains of SecA
E400R/N629D
mutant for analyzing the interaction and regulatory domains of SecA
E635C
mutant for analyzing the interaction and regulatory domains of SecA
Ile3A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
K108R
mutant, defective in ATP binding and protein translocation in vitro, as well as biologically inactive in vivo
Leu2A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
Leu5A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
Leu6A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
N95
truncated version of Escherichia coli SecA, the last 70 residues are lacking
N95CC
two cysteines are introduced into a truncated version of Escherichia coli SecA, at position 636 and 801, the last 70 residues are lacking, mutant is dimeric and fully functional
Phe10A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
R400R/M607T
mutant for analyzing the interaction and regulatory domains of SecA
R642C
mutant for analyzing the interaction and regulatory domains of SecA
R642E
mutant for analyzing the interaction and regulatory domains of SecA
R642E/A628T
mutant for analyzing the interaction and regulatory domains of SecA
R642E/E619K
mutant for analyzing the interaction and regulatory domains of SecA
R642E/H620P
mutant for analyzing the interaction and regulatory domains of SecA
R642E/I627T
mutant for analyzing the interaction and regulatory domains of SecA
R642E/L610P
mutant for analyzing the interaction and regulatory domains of SecA
R642E/M607T
mutant for analyzing the interaction and regulatory domains of SecA
R642E/N629D
mutant for analyzing the interaction and regulatory domains of SecA
R656C
mutant for analyzing the interaction and regulatory domains of SecA
SecADELTA11/N95
monomeric SecA derivative mutant, which lacks extreme terminal residues and shows in vitro and in vivo activities
V9A/F10A
mutation enhances dissociation by 8fold with respect to that of wild-type SecA
Val9A
mutation completely blocks dimerization of SecA in 300 mM KCl buffer
D209A
-
a dominant-negative mutant, binds ATP but is unable to hydrolyze it and is inactive in proOmpA translocation. Mutant generates a translocation intermediate of 18 kDa. Further addition of wild-type SecA causes its translocation into either mature OmpA or another intermediate of 28 kDa that can be translocated into mature by a proton motive force. The addition of excess D209N SecA during translocation causes a topology inversion of SecG
E20D
mutant is not affected by the bulk pH in the range tested, no dramatic change in IC50 value for peptides Gly-Lys, beta-Ala-Lys
E20Q
mutant is not affected by the bulk pH in the range tested, no dramatic change in IC50 value for peptides Gly-Lys, beta-Ala-Lys
E388D
increase in IC50 value for peptides Gly-Lys, beta-Ala-Lys, increase in pH-optimum
E388Q
increase in IC50 value for peptides Gly-Lys, beta-Ala-Lys, increase in pH-optimum
Y477W
-
KM-value for ATP is 1.75fold higher than wild-type value. kcat for ATP is 4.3fold higher than wild-type value
additional information
additional information
a collection of 63 monocysteine mutants for the 901-aminoacid-residue SecA protein is generated for topological analysis of the protein
additional information
-
a collection of 63 monocysteine mutants for the 901-aminoacid-residue SecA protein is generated for topological analysis of the protein
additional information
-
SecY-SecA interactions are studied by an in vivo site-directed cross-linking technique
additional information
-
analysis of the function of the two-helix finger of the SecA ATPase via point mutations
additional information
screening of a library of mutants to identify amino acids critical for peptide transport and identification of 35 single point mutations that result in a full or partial loss of transport activity
additional information
-
screening of a library of mutants to identify amino acids critical for peptide transport and identification of 35 single point mutations that result in a full or partial loss of transport activity
additional information
-
construction of SecA N-terminal deletion mutants. The first small helix, the linker and part of the second helix (residues 2-22) are dispensable for SecA activity in complementing the growth of a SecA temperature-sensitive mutant. Deletions of N-terminal aminoacyl residues 23-25 result in severe progressive retardation of growth. A decrease of SecA activity caused by N-terminal deletions correlates to the loss of SecA membrane binding, formation of lipid-specific domains and channel activity. The N-terminal aminoacyl residues 23-25 play a critical role for SecA binding to membranes and the N-terminal limit of SecA for activity is at the 25th amino acid
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
SecYEG binding stabilizes a cold sodium dodecylsulfate-resistant dimeric state of SecA
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, the ATPase domain is indefinitly stable in phosphate or Tris buffer, pH 8, in the absence of ATP concentrations of up to 25 mg/ml
-
4°C, the ATPase domain is stable for at least 2 days in phosphate or Tris buffer, pH 8, in the absence of ATP concentrations of up to 25 mg/ml
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
using a Q-Sepharose 26/10 anion-exchange column, a SP-Sepharose 16/5 cation exchange column, and a Superose 12 HR gel filtration column
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of a truncated form of SecA lacking the C-terminal inhibitory domain
into the pET5a vector for expression in Escherichia coli BL21DE3 cells
plasmid pT7secA-Cys-0, a derivative of pT7secA2 that has all four cysteine codons within secA changed to serine, is used to create the monocysteine secA mutants
expressed in Escherichia coli after stimulation with 1 mM isopropyl-beta-D-thiogalactoside
-
expressed in Escherichia coli strain BL21(DE3) harboring a special plasmid
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wandersman, C.
Protein and peptide secretion by ABC exporters
Res. Microbiol.
149
163-170
1998
Escherichia coli, Enterococcus faecalis, Gram-negative bacteria, Homo sapiens, Mus sp., Rattus norvegicus
Koronakis, V.; Hughes, C.
Bacterial signal peptide-independent protein export: HlyB-directed secretion of hemolysin
Semin. Cell Biol.
4
7-15
1993
Escherichia coli
Binet, R.; Letoffe, S.; Ghigo, J.M.; Delepelaire, P.; Wandersman, C.
Protein secretion by Gram-negative bacterial ABC exporters - a review
Gene
192
7-11
1997
Escherichia coli, Dickeya chrysanthemi, Gram-negative bacteria, Serratia marcescens
Kranitz, L.; Benabdelhak, H.; Horn, C.; Blight, M.A.; Holland, I.B.; Schmitt, L.
Crystallization and preliminary X-ray analysis of the ATP-binding domain of the ABC transporter haemolsin B from Escherichia coli
Acta Crystallogr. Sect. D
58
539-541
2002
Escherichia coli
Schmitt, L.; Benabdelhak, H.; Blight, M.; Holland, I.B.; Stubbs, M.T.
Crystal structure of the nucleotide-binding domain of the ABC-transporter haemolysinB: Identification of a variable region within ABC helical domains
J. Mol. Biol.
330
333-342
2003
Escherichia coli
Benabdelhak, H.; Schmitt, L.; Horn, C.; Jumel, K.; Blight, M.A.; Holland, I.B.
Positive co-operative activity and dimerization of the isolated ABC ATPase domain of HlyB from Escherichia coli
Biochem. J.
386
489-495
2005
Escherichia coli
Zaitseva, J.; Jenewein, S.; Wiedenmann, A.; Benabdelhak, H.; Holland, I.B.; Schmitt, L.
Functional characterization and ATP-induced dimerization of the isolated ABC-domain of the haemolysin B transporter
Biochemistry
44
9680-9690
2005
Escherichia coli
Nakatogawa, H.; Murakami, A.; Mori, H.; Ito, K.
SecM facilitates translocase function of SecA by localizing its biosynthesis
Genes Dev.
19
436-444
2005
Escherichia coli
de Keyzer, J.; van der Sluis, E.O.; Spelbrink, R.E.; Nijstad, N.; de Kruijff, B.; Nouwen, N.; van der Does, C.; Driessen, A.J.
Covalently dimerized SecA is functional in protein translocation
J. Biol. Chem.
280
35255-35260
2005
Escherichia coli
Deitermann, S.; Sprie, G.S.; Koch, H.G.
A dual function for SecA in the assembly of single spanning membrane proteins in Escherichia coli
J. Biol. Chem.
280
39077-39085
2005
Escherichia coli
Papanikou, E.; Karamanou, S.; Baud, C.; Frank, M.; Sianidis, G.; Keramisanou, D.; Kalodimos, C.G.; Kuhn, A.; Economou, A.
Identification of the preprotein binding domain of SecA
J. Biol. Chem.
280
43209-43217
2005
Escherichia coli
Or, E.; Boyd, D.; Gon, S.; Beckwith, J.; Rapoport, T.
The bacterial ATPase SecA functions as a monomer in protein translocation
J. Biol. Chem.
280
9097-9105
2005
Escherichia coli
Tomkiewicz, D.; Nouwen, N.; van Leeuwen, R.; Tans, S.; Driessen, A.J.
SecA supports a constant rate of preprotein translocation
J. Biol. Chem.
281
15709-15713
2006
Escherichia coli
Jilaveanu, L.B.; Zito, C.R.; Oliver, D.
Dimeric SecA is essential for protein translocation
Proc. Natl. Acad. Sci. USA
102
7511-7516
2005
Escherichia coli
Patel, C.N.; Smith, V.F.; Randall, L.L.
Characterization of three areas of interactions stabilizing complexes between SecA and SecB, two proteins involved in protein export
Protein Sci.
15
1379-1386
2006
Escherichia coli
Or, E.; Rapoport, T.
Cross-linked SecA dimers are not functional in protein translocation
FEBS Lett.
581
2616-2620
2007
Escherichia coli (P10408)
Wang, H.; Na, B.; Yang, H.; Tai, P.C.
Additional in vitro and in vivo evidence for SecA functioning as dimers in the membrane: dissociation into monomers is not essential for protein translocation in Escherichia coli
J. Bacteriol.
190
1413-1418
2008
Escherichia coli (P10408)
Hou, J.M.; DLima, N.G.; Rigel, N.W.; Gibbons, H.S.; McCann, J.R.; Braunstein, M.; Teschke, C.M.
ATPase activity of Mycobacterium tuberculosis SecA1 and SecA2 proteins and its importance to SecA2 function in macrophages
J. Bacteriol.
190
4880-4887
2008
Escherichia coli (P10408), Escherichia coli, Mycobacterium tuberculosis (P9WGP3), Mycobacterium tuberculosis (P9WGP5), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WGP3), Mycobacterium tuberculosis H37Rv (P9WGP5)
Mori, H.; Ito, K.
The long alpha-helix of SecA is important for the ATPase coupling of translocation
J. Biol. Chem.
281
36249-36256
2006
Escherichia coli (P10408)
Sugai, R.; Takemae, K.; Tokuda, H.; Nishiyama, K.
Topology inversion of SecG is essential for cytosolic SecA-dependent stimulation of protein translocation
J. Biol. Chem.
282
29540-29548
2007
Escherichia coli (P10408)
Jilaveanu, L.B.; Oliver, D.B.
In vivo membrane topology of Escherichia coli SecA ATPase reveals extensive periplasmic exposure of multiple functionally important domains clustering on one face of SecA
J. Biol. Chem.
282
4661-4668
2007
Escherichia coli (P10408), Escherichia coli
Mori Hiroyuk, M.H.; Ito Koreak, I.K.
Different modes of SecY-SecA interactions revealed by site-directed in vivo photo-cross-linking
Proc. Natl. Acad. Sci. USA
103
16159-16164
2006
Escherichia coli
Ahn, T.; Yun, C.H.
Ca(2+)-induced stimulation of the membrane binding of Escherichia coli SecA and its association with signal peptides of secretory proteins
Arch. Biochem. Biophys.
486
125-131
2009
Escherichia coli
Robson, A.; Carr, B.; Sessions, R.B.; Collinson, I.
Synthetic peptides identify a second periplasmic site for the plug of the SecYEG protein translocation complex
FEBS Lett.
583
207-212
2009
Escherichia coli
Das, S.; Stivison, E.; Folta-Stogniew, E.; Oliver, D.
Reexamination of the role of the amino terminus of SecA in promoting its dimerization and functional state
J. Bacteriol.
190
7302-7307
2008
Escherichia coli
Mao, C.; Hardy, S.J.; Randall, L.L.
Maximal efficiency of coupling between ATP hydrolysis and translocation of polypeptides mediated by SecB requires two protomers of SecA
J. Bacteriol.
191
978-984
2009
Escherichia coli
Cooper, D.B.; Smith, V.F.; Crane, J.M.; Roth, H.C.; Lilly, A.A.; Randall, L.L.
SecA, the motor of the secretion machine, binds diverse partners on one interactive surface
J. Mol. Biol.
382
74-87
2008
Escherichia coli (P28366)
Nouwen, N.; Berrelkamp, G.; Driessen, A.J.
Charged amino acids in a preprotein inhibit SecA-dependent protein translocation
J. Mol. Biol.
386
1000-1010
2009
Escherichia coli
Erlandson, K.J.; Miller, S.B.; Nam, Y.; Osborne, A.R.; Zimmer, J.; Rapoport, T.A.
A role for the two-helix finger of the SecA ATPase in protein translocation
Nature
455
984-987
2008
Escherichia coli
Chen, W.; Huang, Y.; Reddy Gundala, S.; Yang, H.; Li, N.; Tai, P.C.; Wang, B.
The first low microM SecA inhibitors
Bioorg. Med. Chem.
18
1617-1625
2010
Escherichia coli
Huang, Y.J.; Wang, H.; Gao, F.B.; Li, M.; Yang, H.; Wang, B.; Tai, P.C.
Fluorescein analogues inhibit SecA ATPase: the first sub-micromolar inhibitor of bacterial protein translocation
ChemMedChem
7
571-577
2012
Escherichia coli
Malle, E.; Zhou, H.; Neuhold, J.; Spitzenberger, B.; Klepsch, F.; Pollak, T.; Bergner, O.; Ecker, G.; Stolt-Bergner, P.
Random mutagenesis of the prokaryotic peptide transporter YdgR identifies potential periplasmic gating residues
J. Biol. Chem.
286
23121-23131
2011
Escherichia coli (P77304), Escherichia coli
Morita, K.; Tokuda, H.; Nishiyama, K.
Multiple SecA molecules drive protein translocation across a single translocon with SecG inversion
J. Biol. Chem.
287
455-464
2012
Escherichia coli
Wu, Z.C.; de Keyzer, J.; Kedrov, A.; Driessen, A.J.
Competitive binding of the SecA ATPase and ribosomes to the SecYEG translocon
J. Biol. Chem.
287
7885-7895
2012
Escherichia coli, Escherichia coli SF100
Dalal, K.; Chan, C.S.; Sligar, S.G.; Duong, F.
Two copies of the SecY channel and acidic lipids are necessary to activate the SecA translocation ATPase
Proc. Natl. Acad. Sci. USA
109
4104-4109
2012
Escherichia coli
Jensen, J.; Ernst, H.; Wang, X.; Hald, H.; Ditta, A.; Ismat, F.; Rahman, M.; Mirza, O.
Functional investigation of conserved membrane-embedded glutamate residues in the proton-coupled peptide transporter YjdL
Protein Pept. Lett.
19
282-287
2012
Escherichia coli (P39276), Escherichia coli
Floyd, J.H.; You, Z.; Hsieh, Y.H.; Ma, Y.; Yang, H.; Tai, P.C.
The dispensability and requirement of SecA N-terminal aminoacyl residues for complementation, membrane binding, lipid-specific domains and channel activities
Biochem. Biophys. Res. Commun.
453
138-142
2014
Escherichia coli
Chaudhary, A.S.; Jin, J.; Chen, W.; Tai, P.C.; Wang, B.
Design, syntheses and evaluation of 4-oxo-5-cyano thiouracils as SecA inhibitors
Bioorg. Med. Chem.
23
105-117
2015
Escherichia coli (P10408)
EXTERNAL LINKS (specific for EC-Number 7.4.2.5)
Transporter Classification Database (TCDB): 3.A.1.109.8
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