BRENDA - Enzyme Database

A new paradigm for electron transfer through Escherichia coli nitrate reductase A

Fedor, J.; Rothery, R.; Weiner, J.; Biochemistry 53, 4549-4556 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.7.5.1
gene cluster NarGHI, overexpression in Escherichia coli strain LCB79; gene cluster NarGHI, overexpression in Escherichia coli strain LCB79; gene cluster NarGHI, overexpression in Escherichia coli strain LCB79
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.7.5.1
G65A
site-directed mutageness of subunit NarI, mutant G65A is able to support growth and retains significant quinol:nitrate oxidoreductase activity; site-directed mutageness of subunit NarI, mutant G65A is able to support growth and retains significant quinol:nitrate oxidoreductase activity; site-directed mutageness of subunit NarI, mutant G65A is able to support growth and retains significant quinol:nitrate oxidoreductase activity
Escherichia coli
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.7.5.1
membrane
the NarI subunit anchors the NarGH subunits to the inside of the cytoplasmic membrane; the NarI subunit anchors the NarGH subunits to the inside of the cytoplasmic membrane; the NarI subunit anchors the NarGH subunits to the inside of the cytoplasmic membrane
Escherichia coli
16020
-
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.7.5.1
nitrite + a quinone + H2O
Escherichia coli
-
nitrate + a quinol
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.7.5.1
Escherichia coli
P09152
NarG
-
1.7.5.1
Escherichia coli
P11349
NarH
-
1.7.5.1
Escherichia coli
P11350
NarI
-
Reaction
EC Number
Reaction
Commentary
Organism
1.7.5.1
nitrate + a quinol = nitrite + a quinone + H2O
electrons flow in the overall thermodynamically downhill direction from menaquinol (MQ) or ubiquinol (UQ) through the two hemes of NarI, the four [Fe-S] clusters of NarH, and then through the single [4Fe-4S] cluster of NarG to the Mo-bisPGD cofactor, where nitrate is reduced to nitrite; electrons flow in the overall thermodynamically downhill direction from menaquinol (MQ) or ubiquinol (UQ) through the two hemes of NarI, the four [Fe-S] clusters of NarH, and then through the single [4Fe-4S] cluster of NarG to the Mo-bisPGD cofactor, where nitrate is reduced to nitrite; electrons flow in the overall thermodynamically downhill direction from menaquinol (MQ) or ubiquinol (UQ) through the two hemes of NarI, the four [Fe-S] clusters of NarH, and then through the single [4Fe-4S] cluster of NarG to the Mo-bisPGD cofactor, where nitrate is reduced to nitrite
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.7.5.1
nitrite + a quinone + H2O
-
741900
Escherichia coli
nitrate + a quinol
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.7.5.1
?
x * 140000, NarG subunit, SDS-PAGE; x * 26000, NarI subunit, SDS-PAGE; x * 58000, NarH subunit, SDS-PAGE
Escherichia coli
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.7.5.1
25
-
assay at; assay at; assay at
Escherichia coli
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.7.5.1
7
-
assay at; assay at; assay at
Escherichia coli
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.7.5.1
heme
the membrane subunit (NarI) of Escherichia coli nitrate reductase A (NarGHI) contains two b-type hemes, both of which are the highly anisotropic low-spin type. Heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate; the membrane subunit (NarI) of Escherichia coli nitrate reductase A (NarGHI) contains two b-type hemes, both of which are the highly anisotropic low-spin type. Heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate; the membrane subunit (NarI) of Escherichia coli nitrate reductase A (NarGHI) contains two b-type hemes, both of which are the highly anisotropic low-spin type. Heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate
Escherichia coli
1.7.5.1
molybdo-bis(pyranopterin guanine dinucleotide)
Mo-bisPGD cofactor, bound to subunit NarG. NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively; Mo-bisPGD cofactor, bound to subunit NarG. NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively; Mo-bisPGD cofactor, bound to subunit NarG. NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively
Escherichia coli
1.7.5.1
quinone
heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate; heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate; heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate
Escherichia coli
1.7.5.1
[4Fe-4S]-center
a single tetranuclear iron-sulfur [4Fe-4S] cluster, known as FS0, is bound to subunit NarG. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4); a single tetranuclear iron-sulfur [4Fe-4S] cluster, known as FS0, is bound to subunit NarG. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4); a single tetranuclear iron-sulfur [4Fe-4S] cluster, known as FS0, is bound to subunit NarG. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4)
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.7.5.1
gene cluster NarGHI, overexpression in Escherichia coli strain LCB79
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.7.5.1
heme
the membrane subunit (NarI) of Escherichia coli nitrate reductase A (NarGHI) contains two b-type hemes, both of which are the highly anisotropic low-spin type. Heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate
Escherichia coli
1.7.5.1
molybdo-bis(pyranopterin guanine dinucleotide)
Mo-bisPGD cofactor, bound to subunit NarG. NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively
Escherichia coli
1.7.5.1
quinone
heme bD is distal to NarGH and constitutes part of the quinone binding and oxidation site (Q-site) through the axially coordinating His66 residue and one of the heme bD propionate groups. Bound quinone participates in hydrogen bonds with both the imidazole of His66 and the heme propionate
Escherichia coli
1.7.5.1
[4Fe-4S]-center
a single tetranuclear iron-sulfur [4Fe-4S] cluster, known as FS0, is bound to subunit NarG. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4)
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.7.5.1
G65A
site-directed mutageness of subunit NarI, mutant G65A is able to support growth and retains significant quinol:nitrate oxidoreductase activity
Escherichia coli
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.7.5.1
membrane
the NarI subunit anchors the NarGH subunits to the inside of the cytoplasmic membrane
Escherichia coli
16020
-
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.7.5.1
nitrite + a quinone + H2O
Escherichia coli
-
nitrate + a quinol
-
-
?
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.7.5.1
nitrite + a quinone + H2O
-
741900
Escherichia coli
nitrate + a quinol
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.7.5.1
?
x * 140000, NarG subunit, SDS-PAGE
Escherichia coli
1.7.5.1
?
x * 58000, NarH subunit, SDS-PAGE
Escherichia coli
1.7.5.1
?
x * 26000, NarI subunit, SDS-PAGE
Escherichia coli
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.7.5.1
25
-
assay at
Escherichia coli
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.7.5.1
7
-
assay at
Escherichia coli
General Information
EC Number
General Information
Commentary
Organism
1.7.5.1
malfunction
quinone site variants Lys86 and Gly65, Q-site inhibitor HOQNO, and their effects on heme bD, overview; quinone site variants Lys86 and Gly65, Q-site inhibitor HOQNO, and their effects on heme bD, overview; quinone site variants Lys86 and Gly65, Q-site inhibitor HOQNO, and their effects on heme bD, overview
Escherichia coli
1.7.5.1
additional information
NarGHI comprises a catalytic subunit (NarG, 140 kDa), an electron-transfer subunit (NarH, 58 kDa), and a membrane anchor subunit (NarI, 26 kDa). NarG contains a Mo-bisPGD cofactor that is the site of nitrate reduction as well as a single tetranuclear iron-sulfur ([4Fe-4S]) cluster known as FS0. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4). NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively; NarGHI comprises a catalytic subunit (NarG, 140 kDa), an electron-transfer subunit (NarH, 58 kDa), and a membrane anchor subunit (NarI, 26 kDa). NarG contains a Mo-bisPGD cofactor that is the site of nitrate reduction as well as a single tetranuclear iron-sulfur ([4Fe-4S]) cluster known as FS0. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4). NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively; NarGHI comprises a catalytic subunit (NarG, 140 kDa), an electron-transfer subunit (NarH, 58 kDa), and a membrane anchor subunit (NarI, 26 kDa). NarG contains a Mo-bisPGD cofactor that is the site of nitrate reduction as well as a single tetranuclear iron-sulfur ([4Fe-4S]) cluster known as FS0. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4). NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively
Escherichia coli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.7.5.1
malfunction
quinone site variants Lys86 and Gly65, Q-site inhibitor HOQNO, and their effects on heme bD, overview
Escherichia coli
1.7.5.1
additional information
NarGHI comprises a catalytic subunit (NarG, 140 kDa), an electron-transfer subunit (NarH, 58 kDa), and a membrane anchor subunit (NarI, 26 kDa). NarG contains a Mo-bisPGD cofactor that is the site of nitrate reduction as well as a single tetranuclear iron-sulfur ([4Fe-4S]) cluster known as FS0. NarH contains three [4Fe-4S] clusters (FS1-FS3) and one trinuclear iron-sulfur cluster ([3Fe-4S], FS4). NarI anchors the NarGH subunits to the inside of the cytoplasmic membrane and contains two hemes b that are proximal (bP) and distal (bD) to the NarGH subunits, respectively
Escherichia coli