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Literature summary for 2.7.1.100 extracted from

  • Ku, S.Y.; Yip, P.; Cornell, K.A.; Riscoe, M.K.; Howell, P.L.
    Crystallization and preliminary X-ray analysis of 5'-methylthioribose kinase from Bacillus subtilis and Arabidopsis thaliana (2004), Acta Crystallogr. Sect. D, 60, 116-119.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

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

Metals/Ions Comment Organism Structure
Mg2+
-
Bacillus subtilis
Mg2+
-
Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Arabidopsis thaliana
-
-
-
Bacillus subtilis
-
-
-

Purification (Commentary)

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 (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
MTR kinase
-
Bacillus subtilis
MTR kinase
-
Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Bacillus subtilis
25
-
assay at Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Bacillus subtilis
8
-
assay at Arabidopsis thaliana

Cofactor

Cofactor Comment Organism Structure
ATP
-
Bacillus subtilis
ATP
-
Arabidopsis thaliana