Crystallization (Comment) | Organism |
---|---|
crystallization of purified recombinant enzyme mutants H112S, H113S, and C181S, free, mutant H112S in 0.1 M MES, pH 6.0, 0.2 M sodium acetate, 3 mM sodium azide, or complexed with substrate GTP, mutants H112S and C181S in 0.1 M MOPS, pH 7.0, 10% w/v PEG 6000, 0.1 M ammonium sulfate, or mutant H113S in 0.1 M Tris-HCl, pH 8.5, 0.2 M (NH4)H2PO4, 50% v/v 2-methyl-2,4-pentanediol, addition of GTP for the complex formation, hanging drop vapour diffusion method at room temperature, X-ray diffraction structure determination and analysis at about 2.1-3.2 A resolution, modeling | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C110S | highly reduced activity in both reaction steps using GTP or 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate as substrate, reduced zinc content compared to the wild-type enzyme | Escherichia coli |
C181S | determination of crystal structure, highly reduced activity in both reaction steps using GTP or 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate as substrate, reduced zinc content compared to the wild-type enzyme | Escherichia coli |
H112S | determination of crystal structure, highly reduced activity in both reaction steps using GTP or 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate as substrate, reduced zinc content compared to the wild-type enzyme | Escherichia coli |
H113S | determination of crystal structure, highly reduced activity in both reaction steps using GTP or 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate as substrate, reduced zinc content compared to the wild-type enzyme | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | required, wild-type enzyme contains 0.9 mol of Zn2+ per mol of enzyme subunit, while the zinc content of the mutant enzymes is reduced to below 0.2 Zn2+ per mol of enzyme subunit | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | Escherichia coli | - |
formate + 2-amino-4-hydroxy-6-(erythro-1,2,3-trihydroxypropyl)-dihydropteridine triphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6T5 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
GTP + H2O = formate + 7,8-dihydroneopterin 3'-triphosphate | reaction mechanism, active site structure, GTP binding strcuture | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0001 | - |
below, mutant enzymes, substrate 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate | Escherichia coli |
0.0001 | - |
below, mutant enzymes, substrate GTP | Escherichia coli |
0.088 | - |
wild-type enzyme, substrate 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate | Escherichia coli |
0.091 | - |
wild-type enzyme, substrate GTP | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate + H2O | substrate is the reaction intermediate of the overall reaction | Escherichia coli | formate + 2-amino-4-hydroxy-6-(erythro-1,2,3-trihydroxypropyl)-dihydropteridine triphosphate | - |
? | |
GTP + H2O | - |
Escherichia coli | formate + 2-amino-4-hydroxy-6-(erythro-1,2,3-trihydroxypropyl)-dihydropteridine triphosphate | - |
? | |
GTP + H2O | cyclization to dihydroneopterin triphosphate | Escherichia coli | formate + 2-amino-4-hydroxy-6-(erythro-1,2,3-trihydroxypropyl)-dihydropteridine triphosphate | i.e. dihydroneopterin triphosphate | ? |