Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.4.16 extracted from

  • McCulloch, K.M.; Kinsland, C.; Begley, T.P.; Ealick, S.E.
    Structural studies of thiamin monophosphate kinase in complex with substrates and products (2008), Biochemistry, 47, 3810-3821.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Aquifex aeolicus

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Metals/Ions Comment Organism Structure
Mg2+
-
Aquifex aeolicus

Natural Substrates/ Products (Substrates)

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
-
?

Organism

Organism UniProt Comment Textmining
Aquifex aeolicus O67883
-
-

Reaction

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 and Products (Substrate)

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
-
?

Synonyms

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

Cofactor Comment Organism Structure
ATP
-
Aquifex aeolicus