Cloned (Comment) | Organism |
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expression of C-terminally His6-tagged THI20 in Escherichia coli strain BL21 (DE3) | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
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purified recombinant C-terminally His6-tagged THI20, hanging drop vapour diffusion method, 10 mg/ml protein in 10 mM Tris buffer, pH 7.6, with 30 mM NaCl, is mixed with 10-14% PEG 8000, 0.2 M calcium acetate, and 0.1 M imidazole, pH 7.5, microseeding, 2-3 days, X-ray diffraction structure determination and analysis at 2.68 A resolution, molecular replacement, modelling | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Saccharomyces cerevisiae | THI20 is a trifunctional enzyme that is composed of a ThiD-like N-terminal domain that catalyzes the phosphorylation of 4-amino-5-hydroxymethyl-2-methylpyrimidine to 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate and of 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate to 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate, and a TenA-like C-terminal domain with thiaminase activity | ? | - |
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Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged THI20 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and gel filtration | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | THI20 is a trifunctional enzyme that is composed of a ThiD-like N-terminal domain that catalyzes the phosphorylation of 4-amino-5-hydroxymethyl-2-methylpyrimidine to 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate and of 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate to 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate, and a TenA-like C-terminal domain with thiaminase activity | Saccharomyces cerevisiae | ? | - |
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additional information | thiamin-degradation products are hydrolyzed by thiaminase II yielding 4-amino-5-hydroxymethyl-2-methylpyrimidine. HMP. The C-terminal domain is homologous to Bacillus subtilis TenA, which catalyzes the production of HMP from thiamin degradation products in the salvage pathway | Saccharomyces cerevisiae | ? | - |
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Subunits | Comment | Organism |
---|---|---|
dimer | overall dimeric organization in which N-terminal domains and C-terminal domains form ThiD-like and TenA-like local dimers, A relatively flexible linker region composed of a loop and a short helix joins the two domains, domain organization, overview | Saccharomyces cerevisiae |
More | THI20 is a trifunctional enzyme containing an N-terminal HMP kinase/HMP-P kinase (ThiD-like) domain and a C-terminal thiaminase II (TenA-like) domain. The ThiD-like dimer observed in THI20 resembles other ThiD structures, the TenA-like domain, which is tetrameric in all previously reported structures, forms a dimer | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
Thi20 | - |
Saccharomyces cerevisiae |
thiaminase II | - |
Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
metabolism | thiamin-degradation products are hydrolyzed by thiaminase II, yielding 4-amino-5-hydroxymethyl-2-methylpyrimidine. This compound is an intermediate in thiamin biosynthesis. The enzyme is involved in the biosynthesis of thiamin diphosphate, the active form of the cofactor | Saccharomyces cerevisiae |
additional information | THI20 is a trifunctional enzyme containing an N-terminal HMP kinase/HMP-P kinase (ThiD-like) domain and a C-terminal thiaminase II (TenA-like) domain | Saccharomyces cerevisiae |