Cloned (Comment) | Organism |
---|---|
expression as His-tagged and non-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
expression as His-tagged enzyme in Escherichia coli strain BL21(DE3) in inclusion bodies | Arabidopsis thaliana |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, 0.002 ml of protein solution containing 10 mg/ml protein in 20 mM imidazole, pH 7.6, 2 mM DTT, 1 mM 5-methyl-5-thio-D-ribose and 1 mM ATP-gamma-S, is mixed an equal volume of reservoir solution containing 10% w/v PEG 8000, 0.1 M sodium cacodylate, pH 6.5, and 0.2 M magnesium acetate, equilibration against 0.5 ml reservoir solution, room temperature, 3 days, cryoprotection by soaking in reservoir solution plus 25% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 1.9 A resolution, identification of heavy-atom derivatives | Arabidopsis thaliana |
purified recombinant non-tagged enzyme, hanging drop vapour diffusion method, 0.002-0.003 ml of protein solution containing 7.5-15 mg/ml protein in 10 mM potassium phosphate, pH 7.4, 2 mM DTT, 1 mM 5-methyl-5-thio-D-ribose and 1 mM ATP-gamma-S, or 8 mM CHAPS, is mixed an equal volume of reservoir solution containing 22% w/v PEG 2000 monomethylether, 0.1 M Tris-HCl, pH 7.5, and 0.3 M sodium acetate, equilibration against 0.5 ml reservoir solution, room temperature, 3 days, cryoprotection by soaking in reservoir solution plus 25% v/v ethylene glycol, X-ray diffraction structure determination and analysis at 3.0 A resolution without detergent, and at 2.2 A resolution in presence of CHAPS, no crystallization of His-tagged enzyme possible, identification of heavy-atom derivatives | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Bacillus subtilis | |
Mg2+ | - |
Arabidopsis thaliana |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + S-methyl-5-thio-D-ribose | Bacillus subtilis | - |
ADP + S-methyl-5-thio-D-ribose 1-phosphate | - |
? | |
ATP + S-methyl-5-thio-D-ribose | Arabidopsis thaliana | - |
ADP + S-methyl-5-thio-D-ribose 1-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
- |
- |
Bacillus subtilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged and non-tagged enzymes from Escherichia coli strain BL21(DE3), the latter by anion exchange chromatography, ultracentrifucation and gel filtration | Bacillus subtilis |
recombinant refolded His-tagged enzyme from Escherichia coli by centrifugation, nickel affinity chromatography, ultrafiltration and gel filtration | Arabidopsis thaliana |
Renatured (Comment) | Organism |
---|---|
solubilization and refolding of His-tagged enzyme from Escherichia coli inclusion bodies by 6 M guanidium hydrochloride, 0.7 M NaCl, and 20 mM imidazole, at pH 8.0, 30 min at 4°C | Arabidopsis thaliana |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1 | - |
purified recombinant His-tagged enzyme | Arabidopsis thaliana |
1.5 | - |
purified recombinant non-tagged enzyme | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + S-methyl-5-thio-D-ribose | - |
Bacillus subtilis | ADP + S-methyl-5-thio-D-ribose 1-phosphate | - |
? | |
ATP + S-methyl-5-thio-D-ribose | - |
Arabidopsis thaliana | ADP + S-methyl-5-thio-D-ribose 1-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MTR kinase | - |
Bacillus subtilis |
MTR kinase | - |
Arabidopsis thaliana |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
25 | - |
assay at | Arabidopsis thaliana |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Bacillus subtilis |
8 | - |
assay at | Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Bacillus subtilis | |
ATP | - |
Arabidopsis thaliana |