EC Number | Cloned (Comment) | Organism |
---|---|---|
3.5.4.25 | expressed in Escherichia coli BL21(DE3) cells | Mycobacterium tuberculosis |
4.1.99.12 | recombinant expression of His-tagged DHBPS and GCHII domains of MtbribA2 in Escherichia coli strain BL21 (DE3) | Mycobacterium tuberculosis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.5.4.25 | sitting drop vapor diffusion method, using 0.1 M Na HEPES pH 7.5, 15% (w/v) PEG 8000 | Mycobacterium tuberculosis |
4.1.99.12 | purified recombinant bifunctional enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 20-25 mg/ml protein in 20 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM DTT, with 0.001 ml of reservoir solutioncontaining 0.1 M Na HEPES pH 7.5, 15%(w/v) PEG 8000, equilibration against 0.04 ml reservoir solution, 20°C, 2-4 days, X-ray diffraction structure determination and analysis at 3.0 A resolution | Mycobacterium tuberculosis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.4.25 | 0.1267 | - |
GTP | full-length bifunctional enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis | |
3.5.4.25 | 0.1505 | - |
GTP | GCHII-domain of enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.5.4.25 | Mg2+ | required | Mycobacterium tuberculosis | |
3.5.4.25 | Zn2+ | contains zinc | Mycobacterium tuberculosis | |
4.1.99.12 | Mg2+ | essential for catalytic activity and in stabilization of the substrate | Mycobacterium tuberculosis | |
4.1.99.12 | additional information | the GTP cyclohydrolase II subunit has also bound Zn2+ ions | Mycobacterium tuberculosis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.5.4.25 | 48163 | - |
2 * 48163, MALDI-TOF mass spectrometry | Mycobacterium tuberculosis |
3.5.4.25 | 90000 | - |
gel filtration | Mycobacterium tuberculosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.4.25 | GTP + H2O | Mycobacterium tuberculosis | - |
formate + 2,5-diamino-6-hydroxy-4-(5-phospho-D-ribosylamino)pyrimidine + diphosphate | - |
? | |
3.5.4.25 | GTP + H2O | Mycobacterium tuberculosis H37Rv | - |
formate + 2,5-diamino-6-hydroxy-4-(5-phospho-D-ribosylamino)pyrimidine + diphosphate | - |
? | |
4.1.99.12 | D-ribulose 5-phosphate | Mycobacterium tuberculosis | - |
formate + L-3,4-dihydroxybutan-2-one 4-phosphate | - |
? | |
4.1.99.12 | D-ribulose 5-phosphate | Mycobacterium tuberculosis H37Ra / ATCC 25177 | - |
formate + L-3,4-dihydroxybutan-2-one 4-phosphate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.5.4.25 | Mycobacterium tuberculosis | P9WHF1 | - |
- |
3.5.4.25 | Mycobacterium tuberculosis H37Rv | P9WHF1 | - |
- |
4.1.99.12 | Mycobacterium tuberculosis | A5U2B7 | - |
- |
4.1.99.12 | Mycobacterium tuberculosis H37Ra / ATCC 25177 | A5U2B7 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.5.4.25 | ammonium sulfate precipitation, Ni-NTA column chromatography, and Sephacryl S-200 gel filtration | Mycobacterium tuberculosis |
4.1.99.12 | recombinant His-tagged DHBPS and GCHII domains from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and gel filtration | Mycobacterium tuberculosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.4.25 | GTP + H2O | - |
Mycobacterium tuberculosis | formate + 2,5-diamino-6-hydroxy-4-(5-phospho-D-ribosylamino)pyrimidine + diphosphate | - |
? | |
3.5.4.25 | GTP + H2O | - |
Mycobacterium tuberculosis H37Rv | formate + 2,5-diamino-6-hydroxy-4-(5-phospho-D-ribosylamino)pyrimidine + diphosphate | - |
? | |
4.1.99.12 | D-ribulose 5-phosphate | - |
Mycobacterium tuberculosis | formate + L-3,4-dihydroxybutan-2-one 4-phosphate | - |
? | |
4.1.99.12 | D-ribulose 5-phosphate | - |
Mycobacterium tuberculosis H37Ra / ATCC 25177 | formate + L-3,4-dihydroxybutan-2-one 4-phosphate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.5.4.25 | homodimer | 2 * 48163, MALDI-TOF mass spectrometry | Mycobacterium tuberculosis |
4.1.99.12 | dimer | two conformationally different molecules of Mtb-ribA2 in the asymmetric unit that form a dimer via their GCHII domains. DHBPS and GCHII functional homodimers form a long helical-like oligomer, but the enzyme exists as a dimer in solution, isolated subunit Mtb-GCHII is a dimer while Mtb-DHBPS exists as amonomer in solution. The N-terminal DHBPS domain of Mtb-ribA2 shows an alpha + beta fold consisting of a central eight-stranded beta-sheet (beta1-beta8) surrounded by seven helices (alpha1-7alpha7) | Mycobacterium tuberculosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.5.4.25 | GCHII | - |
Mycobacterium tuberculosis |
3.5.4.25 | ribA2 | bifunctional enzyme with 3,4-dihydroxy-2-butanone 4-phosphate synthase and GTP cyclohydrolase II activities | Mycobacterium tuberculosis |
4.1.99.12 | DHBPS | - |
Mycobacterium tuberculosis |
4.1.99.12 | DHBPS/GCHII | - |
Mycobacterium tuberculosis |
4.1.99.12 | Mtb-ribA2 | - |
Mycobacterium tuberculosis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.4.25 | 0.0033 | - |
GTP | full-length bifunctional enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis | |
3.5.4.25 | 0.005 | - |
GTP | GCHII-domain of enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.1.99.12 | 8 | - |
assay at | Mycobacterium tuberculosis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
4.1.99.12 | additional information | an essential bifunctional 3,4-dihydroxy-2-butanone 4-phosphate synthase/GTP cyclohydrolase II (DHBPS/GCHII) enzyme from Mycobacterium tuberculosis, Mtb-ribA2. The enzyme is composed of two conformationally different molecules of Mtb-ribA2 in the asymmetric unit that form a dimer via their GCHII domains. DHBPS and GCHII functional homodimers form a long helical-like oligomer, but the enzyme exists as a dimer in solution. The DHBPS subunit possesses an active-site loop1, a substrate channelling loop2, and a pH-sensitive loop3, structure comparisons, overview | Mycobacterium tuberculosis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.4.25 | 1.4 | - |
GTP | full-length bifunctional enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis | |
3.5.4.25 | 2.1 | - |
GTP | GCHII-domain of enzyme ribA2, at pH 8.0 and 37°C | Mycobacterium tuberculosis |