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Literature summary extracted from
- Imidazole glycerol phosphate synthase from Thermotoga maritima. Quaternary structure, steady-state kinetics, and reaction mechanism of the bienzyme complex (2001), J. Biol. Chem., 276, 20387-20396 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | 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 |
Protein Variants
| EC Number | Protein Variants | Comment | 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 | Comment | 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 | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 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 | Comment | 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 (Comment) | 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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 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 | 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 | 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 | 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 | 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 | 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 | 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 | - |
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 | 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 | - |
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 | 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 | 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 | 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 | Comment | 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] | Comment | 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 | Comment | 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 | Comment | Organism |
|---|---|---|---|---|
| 4.3.1.B2 | 8.5 | - |
assay at | Thermotoga maritima |
General Information
| EC Number | General Information | Comment | 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 | Comment | 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 | |
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