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Literature summary for 4.1.99.17 extracted from
- Analysis of ThiC variants in the context of the metabolic network of Salmonella enterica (2012), J. Bacteriol., 194, 6088-6095.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 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
| Protein Variants | Comment | Organism |
|---|---|---|
| A527T | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
| D468N | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| D509G | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
| D61N/G513E | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| E281K | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
| G273N | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
| G355D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| G472D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| G479R | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| G481S | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| G486D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| G92D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| H501Y | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| additional information | mutant phenotypes, overview | Salmonella enterica |
| P498L | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
| R397H | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| R544C | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| S247F | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
| V267M | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | the enzyme requires an [Fe-S] cluster | Salmonella enterica |  |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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 | ? | |
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Salmonella enterica | - |
serovar Typhimurium LT2, gene thiC | - |
| Salmonella enterica DM851 | - |
serovar Typhimurium LT2, gene thiC | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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 | - |
? | |
| 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 | ? | |
| 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 | - |
? | |
| 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 | ? | |
| additional information | reaction assay in the anaerobic chamber | Salmonella enterica | ? | - |
? | |
| additional information | reaction assay in the anaerobic chamber | Salmonella enterica DM851 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| HMP-P synthase | - |
Salmonella enterica |
| thiC | - |
Salmonella enterica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Salmonella enterica |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 85 | - |
purified recombinant His-tagged wild-type and mutant enzymes, 3 min, inactivation | Salmonella enterica |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Salmonella enterica |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| S-adenosyl-L-methionine | - |
Salmonella enterica | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme is a member of the radical S-adenosylmethionine (AdoMet) superfamily | Salmonella enterica |
| malfunction | a compromised iron-sulfur ([Fe-S]) cluster metabolism reduces ThiC activity | Salmonella enterica |
| 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 |
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