Literature summary for 2.7.1.50 extracted from

Paul, D.; Chatterjee, A.; Begley, T.P.; Ealick, S.E.
Domain organization in Candida glabrata THI6, a bifunctional enzyme required for thiamin biosynthesis in eukaryotes (2010), Biochemistry, 49, 9922-9934.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	[Candida] glabrata

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged enzyme, in complex with beta,gamma-methyleneadenosine 5'-diphosphate, thiamin phosphate, 4-amino-5-hydroxymethyl-2-trifluoromethylpyrimidine diphosphate, or 4-methyl-5-hydroxyethylthiazole phosphate, hanging-drop vapor diffusion method, mixing of0.001 ml protein solution with 0.001 ml of reservoir solution containing 0.1 M HEPES, pH 7.5, 0.2-0.25 M MgCl2, and 25-32% PEG400, 22°C, 2-3 days, method optimization, X-ray diffraction structure determination and analysis at 2.6-3.3 A resolution, modeling	[Candida] glabrata

Crystallization (Comment)

Organism

purified recombinant detagged enzyme, in complex with beta,gamma-methyleneadenosine 5'-diphosphate, thiamin phosphate, 4-amino-5-hydroxymethyl-2-trifluoromethylpyrimidine diphosphate, or 4-methyl-5-hydroxyethylthiazole phosphate, hanging-drop vapor diffusion method, mixing of0.001 ml protein solution with 0.001 ml of reservoir solution containing 0.1 M HEPES, pH 7.5, 0.2-0.25 M MgCl2, and 25-32% PEG400, 22°C, 2-3 days, method optimization, X-ray diffraction structure determination and analysis at 2.6-3.3 A resolution, modeling

[Candida] glabrata

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required, two ions per enzyme molecule, binding structure involving C467, D340, and E372, overview	[Candida] glabrata

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole	[Candida] glabrata	-	ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole	-	?

Organism

Organism	UniProt	Comment	Textmining
[Candida] glabrata	Q6FV03	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the tag by TEV protease, and another step of nickel affinity chromatography	[Candida] glabrata

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole	-	[Candida] glabrata	ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole	-	?

Subunits

Subunits	Comment	Organism
homohexamer	the six protomers form a cage-like structure. Each protomer is composed of two domains, which are structurally homologous to their monofunctional bacterial counterparts. Two loop regions not found in the bacterial enzymes provide interactions between the two domains	[Candida] glabrata

Synonyms

Synonyms	Comment	Organism
4-methyl-5-hydroxyethylthiazole kinase	C-terminal domain of TH16	[Candida] glabrata
THI6	-	[Candida] glabrata
ThiM	-	[Candida] glabrata

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	[Candida] glabrata

General Information

General Information	Comment	Organism
evolution	THI6 is a bifunctional enzyme found in the thiamin biosynthetic pathway in eukaryotes. In prokaryotes, thiamin phosphate synthase and 4-methyl-5-hydroxyethylthiazole kinase are separate gene products	[Candida] glabrata
metabolism	THI6 is a bifunctional enzyme of the thiamin biosynthetic pathway	[Candida] glabrata
physiological function	the N-terminal domain of THI6 catalyzes the ligation of the thiamin thiazole and pyrimidine moieties to form thiamin phosphate, and the C-terminal domain catalyzes the phosphorylation of 4-methyl-5-hydroxyethylthiazole in a salvage pathway	[Candida] glabrata