Cloned (Comment) | Organism |
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expression in Escherichia coli | Aquifex aeolicus |
Crystallization (Comment) | Organism |
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purified recombinant enzyme in complex with nonhydrolyzable AMP-PCP and TMP, or with the reaction products ADP and TPP, ThiL is incubated with 4 mM AMP-PCP or ATP and 4 mM MgCl2 on ice for 1 h prior to crystallization experiments, optimized conditions for the ThiL binary complexes include 100 mM HEPES, pH 7.4, 6-11% isopropyl alcohol, 200 mM NaCl, and 4-6% 2-methyl-2,4-pentanediol, 100 mM CaCl2 is used as an additive in a 1:9 ratio with the reservoir solution, TMP is soaked into the co-crystallized ThiL and AMP-PCP crystals by transferring the crystals to a solution containing the mother liquor and 20 mM TMP and allowing the crystals to soak for 5 min before being frozen. To obtain ThiL ADP-TPP crystals, ThiL is incubated on ice with 4 mM ADP, 4 mM TPP, and 4 mM MgCl2, optimized conditions for the product complex include 100 mM imidazole, pH 8.0, 7-10% PEG 8000, and 100-250 mM Ca(C2H3O2)2, 3-7 days, cryoprotection by 15% ethylene glycol, X-ray diffraction structure determination and analysis at 1.5-2.6 A resolution | Aquifex aeolicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Aquifex aeolicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + thiamine phosphate | Aquifex aeolicus | ThiL catalyzes the ATP-dependent phosphorylation of thiamin monophosphate to form thiamin diphosphate, the active form of vitamin B1 | ADP + thiamine diphosphate | - |
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Organism | UniProt | Comment | Textmining |
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Aquifex aeolicus | O67883 | - |
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Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + thiamine phosphate = ADP + thiamine diphosphate | Aquifex aeolicus ThiL utilizes a direct, inline transfer of the gamma-phosphate of ATP to TMP rather than a phosphorylated enzyme intermediate, reaction mechanism, overview | Aquifex aeolicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + thiamine phosphate | ThiL catalyzes the ATP-dependent phosphorylation of thiamin monophosphate to form thiamin diphosphate, the active form of vitamin B1 | Aquifex aeolicus | ADP + thiamine diphosphate | - |
? | |
ATP + thiamine phosphate | substrate and product binding site structures, conformational changes in ThiL upon substrate binding, overview | Aquifex aeolicus | ADP + thiamine diphosphate | - |
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Synonyms | Comment | Organism |
---|---|---|
More | ThiL is a member of a small ATP binding superfamily | Aquifex aeolicus |
thiamin monophosphate kinase | - |
Aquifex aeolicus |
ThiL | - |
Aquifex aeolicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Aquifex aeolicus |