BRENDA - Enzyme Database

Imidazole glycerol phosphate synthase from Thermotoga maritima. Quaternary structure, steady-state kinetics, and reaction mechanism of the bienzyme complex

Beismann-Driemeyer, S.; Sterner, R.; J. Biol. Chem. 276, 20387-20396 (2001)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
4.3.1.B2
subunits HisH and HisF from Thermotoga maritima are produced in Escherichia coli
Thermotoga maritima
4.3.2.10
tHisH and tHisF from Thermotoga maritima are produced in Escherichia coli
Thermotoga maritima
Engineering
EC Number
Protein Variants
Commentary
Organism
4.3.1.B2
C9A
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
D11N
mutation in subunit HisF. Catalytic efficiency kcat/Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.1.B2
D130N
the kcat value is reduced by a factor of about 400-500, and the Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is increased almost 20fold. Catalytic efficiency kcat/Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.1.B2
D176N
mutation in subunit HisF. The variant tHisF_D176N shows a 40-50 fold decrease in kcat, both in isolated form and in complex with tHisH
Thermotoga maritima
4.3.1.B2
D183N
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
D51N
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
K19S
mutation in subunit HisF. The ammonia-dependent reactions of isolated subunit HisF_K19S is similarly efficient as that of wild-type HisF. The efficiencies of the glutamine-dependent reactions of the tHisHtHisF_K19S complex are significantly impaired
Thermotoga maritima
4.3.1.B2
additional information
eight conserved amino acids at the putative active site of subunit HisF are exchanged by site-directed mutagenesis, and the purified variants are investigated by steady state kinetics. Aspartate 11 appears to be essential for the synthase activity both in vitro and in vivo, and aspartate 130 can be partially replaced only by glutamate
Thermotoga maritima
4.3.1.B2
N103A
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.2.10
C9A
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_C9A are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
4.3.2.10
D130N
mutant of subunit HisF. The catalytic efficiency kcat/Km for 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.2.10
D176N
mutant of subunit HisF. Variant tHisF_D176N shows a 40-50 fold decrease in kcat, both in isolated form and in complex with tHisH
Thermotoga maritima
4.3.2.10
D183N
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_D183N are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue. Both kcat and Km are drastically impaired in the mutant enzyme
Thermotoga maritima
4.3.2.10
D51N
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_D51N are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
4.3.2.10
K19S
mutant of subunit HisF. The ammonia-dependent reactions of isolated tHisF_K19S are similarly efficient as those of wild-type tHisF. In contrast, the efficiencies of the glutamine-dependent reactions of the tHisHtHisF_K19S complex are significantly impaired
Thermotoga maritima
4.3.2.10
N103A
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_N103A are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.0015
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
0.32
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
0.0015
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
0.0017
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
0.32
-
L-glutamine
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
2.2
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
Thermotoga maritima
the enzyme links histidine and de novo purine biosynthesis
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
Thermotoga maritima ATCC 43589
the enzyme links histidine and de novo purine biosynthesis
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
4.3.1.B2
Thermotoga maritima
Q9X0C8 and Q9X0C6
Q9X0C8: subunit HisH, Q9X0C6: subunit HisF
-
4.3.1.B2
Thermotoga maritima DSM 3109
Q9X0C8 and Q9X0C6
Q9X0C8: subunit HisH, Q9X0C6: subunit HisF
-
4.3.2.10
Thermotoga maritima
Q9X0C6 AND Q9X0C8
Q9X0C6: subunit HisF, Q9X0C8: subunit HisH
-
4.3.2.10
Thermotoga maritima ATCC 43589
Q9X0C6 AND Q9X0C8
Q9X0C6: subunit HisF, Q9X0C8: subunit HisH
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
4.3.1.B2
subunits HisH and HisF from Thermotoga maritima are produced in Escherichia coli and purified. Wild-type and active site mutants
Thermotoga maritima
4.3.2.10
-
Thermotoga maritima
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4.3.1.B2
L-glutamine + H2O
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazoleglycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
L-glutamate + NH3
-
-
-
?
4.3.1.B2
L-glutamine + H2O
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazoleglycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
L-glutamate + NH3
-
-
-
?
4.3.1.B2
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
-
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
the enzyme links histidine and de novo purine biosynthesis
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
-
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
the enzyme links histidine and de novo purine biosynthesis
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+
ammonia-dependent ImGP synthase reaction of isolated HisF subunit
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+
ammonia-dependent ImGP synthase reaction of isolated HisF subunit
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.3.2.10
dimer
imidazole glycerol phosphate synthase constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Isolated tHisH shows no detectable glutaminase activity but is stimulated by complex formation with tHisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
Thermotoga maritima
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.1.B2
25
-
assay at
Thermotoga maritima
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.4
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
0.8
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
0.4
-
L-glutamine
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
0.8
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
2
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
2.2
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.1.B2
8.5
-
assay at
Thermotoga maritima
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.1.B2
subunits HisH and HisF from Thermotoga maritima are produced in Escherichia coli
Thermotoga maritima
4.3.2.10
tHisH and tHisF from Thermotoga maritima are produced in Escherichia coli
Thermotoga maritima
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
4.3.1.B2
C9A
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
D11N
mutation in subunit HisF. Catalytic efficiency kcat/Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.1.B2
D130N
the kcat value is reduced by a factor of about 400-500, and the Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is increased almost 20fold. Catalytic efficiency kcat/Km for N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.1.B2
D176N
mutation in subunit HisF. The variant tHisF_D176N shows a 40-50 fold decrease in kcat, both in isolated form and in complex with tHisH
Thermotoga maritima
4.3.1.B2
D183N
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
D51N
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.1.B2
K19S
mutation in subunit HisF. The ammonia-dependent reactions of isolated subunit HisF_K19S is similarly efficient as that of wild-type HisF. The efficiencies of the glutamine-dependent reactions of the tHisHtHisF_K19S complex are significantly impaired
Thermotoga maritima
4.3.1.B2
additional information
eight conserved amino acids at the putative active site of subunit HisF are exchanged by site-directed mutagenesis, and the purified variants are investigated by steady state kinetics. Aspartate 11 appears to be essential for the synthase activity both in vitro and in vivo, and aspartate 130 can be partially replaced only by glutamate
Thermotoga maritima
4.3.1.B2
N103A
mutation in subunit HisF. Catalytic efficiencies kcat/Km of both isolated and complexed mutant subunit HisF is not significantly different from wild-type HisF
Thermotoga maritima
4.3.2.10
C9A
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_C9A are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
4.3.2.10
D130N
mutant of subunit HisF. The catalytic efficiency kcat/Km for 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide is decreased by approximately 5 orders of magnitude
Thermotoga maritima
4.3.2.10
D176N
mutant of subunit HisF. Variant tHisF_D176N shows a 40-50 fold decrease in kcat, both in isolated form and in complex with tHisH
Thermotoga maritima
4.3.2.10
D183N
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_D183N are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue. Both kcat and Km are drastically impaired in the mutant enzyme
Thermotoga maritima
4.3.2.10
D51N
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_D51N are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
4.3.2.10
K19S
mutant of subunit HisF. The ammonia-dependent reactions of isolated tHisF_K19S are similarly efficient as those of wild-type tHisF. In contrast, the efficiencies of the glutamine-dependent reactions of the tHisHtHisF_K19S complex are significantly impaired
Thermotoga maritima
4.3.2.10
N103A
mutant of subunit HisF. The catalytic efficiencies kcat/Km of both isolated and complexed tHisF_N103A are not significantly different from wild-type tHisF, ruling out any central catalytic role for the replaced residue
Thermotoga maritima
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.0015
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
0.32
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
0.0015
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
0.0017
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
0.32
-
L-glutamine
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
2.2
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
Thermotoga maritima
the enzyme links histidine and de novo purine biosynthesis
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
Thermotoga maritima ATCC 43589
the enzyme links histidine and de novo purine biosynthesis
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.1.B2
subunits HisH and HisF from Thermotoga maritima are produced in Escherichia coli and purified. Wild-type and active site mutants
Thermotoga maritima
4.3.2.10
-
Thermotoga maritima
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4.3.1.B2
L-glutamine + H2O
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazoleglycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
L-glutamate + NH3
-
-
-
?
4.3.1.B2
L-glutamine + H2O
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazoleglycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
L-glutamate + NH3
-
-
-
?
4.3.1.B2
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.1.B2
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3
the enzyme constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Nascent ammonia produced by HisH reacts at the active site of HisF with N'-((5'-phosphoribulosyl)formimino)-5-aminoimidazole-4-carboxamide-ribonucleotide to yield the products imidazole glycerol phosphate and 5-aminoimidazole-4-carboxamide ribotide. Isolated subunit HisH shows no detectable glutaminase activity but is stimulated by complex formation with subunit HisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
748148
Thermotoga maritima DSM 3109
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
-
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
the enzyme links histidine and de novo purine biosynthesis
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
-
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine
the enzyme links histidine and de novo purine biosynthesis
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+
ammonia-dependent ImGP synthase reaction of isolated HisF subunit
748148
Thermotoga maritima
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O
-
-
-
?
4.3.2.10
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+
ammonia-dependent ImGP synthase reaction of isolated HisF subunit
748148
Thermotoga maritima ATCC 43589
5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.2.10
dimer
imidazole glycerol phosphate synthase constitutes a bienzyme complex of the glutaminase subunit HisH and the synthase subunit HisF. Isolated tHisH shows no detectable glutaminase activity but is stimulated by complex formation with tHisF to which either the product imidazole glycerol phosphate or a substrate analogue are bound
Thermotoga maritima
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.3.1.B2
25
-
assay at
Thermotoga maritima
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.4
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
0.8
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
0.4
-
L-glutamine
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
0.8
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
2
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
2.2
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.3.1.B2
8.5
-
assay at
Thermotoga maritima
General Information
EC Number
General Information
Commentary
Organism
4.3.1.B2
metabolism
the enzyme links histidine and de novo purine biosynthesis
Thermotoga maritima
4.3.2.10
metabolism
the enzyme links histidine and de novo purine biosynthesis
Thermotoga maritima
General Information (protein specific)
EC Number
General Information
Commentary
Organism
4.3.1.B2
metabolism
the enzyme links histidine and de novo purine biosynthesis
Thermotoga maritima
4.3.2.10
metabolism
the enzyme links histidine and de novo purine biosynthesis
Thermotoga maritima
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.0006
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
1.4
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
1.4
-
L-glutamine
pH 8.0, 25°C, wild-type enzyme HisHF
Thermotoga maritima
4.3.2.10
600
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
900
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
1300
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.3.1.B2
0.0006
-
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
pH 8.5, 25°C
Thermotoga maritima
4.3.1.B2
1.4
-
L-glutamine
pH 8.5, 25°C
Thermotoga maritima
4.3.2.10
1.4
-
L-glutamine
pH 8.0, 25°C, wild-type enzyme HisHF
Thermotoga maritima
4.3.2.10
600
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.0, 25°C, wild-type HisH-HisF complex
Thermotoga maritima
4.3.2.10
900
-
NH4+
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima
4.3.2.10
1300
-
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide
pH 8.5, 25°C, ammonia-dependent reaction of isolated HisF subunit, wild-type
Thermotoga maritima