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Literature summary extracted from

  • Palmer, L.D.; Dougherty, M.J.; Downs, D.M.
    Analysis of ThiC variants in the context of the metabolic network of Salmonella enterica (2012), J. Bacteriol., 194, 6088-6095.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.99.17 gene thiC, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21AI, which also expresses Azotobacter vinelandii [Fe-S] cluster loading genes (iscSUA, hscBA, fdx, orf3, ndK) Salmonella enterica

Protein Variants

EC Number Protein Variants Comment Organism
4.1.99.17 A527T random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica
4.1.99.17 D468N random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 D509G random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica
4.1.99.17 D61N/G513E random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 E281K random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica
4.1.99.17 G273N random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica
4.1.99.17 G355D random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 G472D random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 G479R random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 G481S random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 G486D random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 G92D random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 H501Y random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 additional information mutant phenotypes, overview Salmonella enterica
4.1.99.17 P498L random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica
4.1.99.17 R397H random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 R544C random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 S247F random mutagenesis by by hydroxylamine, inactive mutant Salmonella enterica
4.1.99.17 V267M random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme Salmonella enterica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.99.17 Fe2+ the enzyme requires an [Fe-S] cluster Salmonella enterica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine Salmonella enterica
-
 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate ?
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine Salmonella enterica DM851
-
 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate ?

Organism

EC Number Organism UniProt Comment Textmining
4.1.99.17 Salmonella enterica
-
 serovar Typhimurium LT2, gene thiC
-
4.1.99.17 Salmonella enterica DM851
-
 serovar Typhimurium LT2, gene thiC
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.99.17 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21AI by nickel affinity chromatography and gel filtration Salmonella enterica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine
-
 Salmonella enterica 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO
-
 ?
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine
-
 Salmonella enterica 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate ?
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine
-
 Salmonella enterica DM851 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO
-
 ?
4.1.99.17 5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine
-
 Salmonella enterica DM851 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate ?
4.1.99.17 additional information reaction assay in the anaerobic chamber Salmonella enterica ?
-
 ?
4.1.99.17 additional information reaction assay in the anaerobic chamber Salmonella enterica DM851 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.1.99.17 HMP-P synthase
-
 Salmonella enterica
4.1.99.17 thiC
-
 Salmonella enterica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.99.17 37
-
 assay at Salmonella enterica

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.99.17 85
-
 purified recombinant His-tagged wild-type and mutant enzymes, 3 min, inactivation Salmonella enterica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.99.17 7.5
-
 assay at Salmonella enterica

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.99.17 S-adenosyl-L-methionine
-
 Salmonella enterica

General Information

EC Number General Information Comment Organism
4.1.99.17 evolution the enzyme is a member of the radical S-adenosylmethionine (AdoMet) superfamily Salmonella enterica
4.1.99.17 malfunction a compromised iron-sulfur ([Fe-S]) cluster metabolism reduces ThiC activity Salmonella enterica
4.1.99.17 metabolism the enzyme catalyzes the biosynthesis of on part of the thiamine diphosphate cofactor that is essentially used by enzymes in central metabolism such as pyruvate dehydrogenase and2-oxoglutarate dehydrogenase to stabilize the acyl carbanion Salmonella enterica