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Literature summary for 2.4.2.19 extracted from
- Interactions at the 2 and 5 positions of 5-phosphoribosyl pyrophosphate are essential in Salmonella typhimurium quinolinate phosphoribosyltransferase (2010), Biochemistry, 49, 1377-1387.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| nadC coding sequence cloned into the pRSETC expression vector, overexpressed in Escherichia coli nadC-deleted host strain ZB100 | Salmonella enterica subsp. enterica serovar Typhimurium |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D235A | does not affect ligand binding or catalysis, KD value for quinolinic acid is similar to wild-type, increases KD value for 5-phospho-alpha-D-ribose 1-diphosphate by 2fold | Salmonella enterica subsp. enterica serovar Typhimurium |
| E214A | increases KD value for quinolinic acid by 2fold and for 5-phospho-alpha-D-ribose 1-diphosphate by 15fold, causes at least a 4000fold reduction in kcat. Presence of benzene-1,2-dicarboxylic acid results in 3.4fold tightening of 5-phospho-alpha-D-ribose 1-diphosphate binding | Salmonella enterica subsp. enterica serovar Typhimurium |
| E214D | KD value for quinolinic acid is similar to wild-type, increases KD value for 5-phospho-alpha-D-ribose 1-diphosphate by 10fold. Presence of benzene-1,2-dicarboxylic acid results in 6fold tightening of 5-phospho-alpha-D-ribose 1-diphosphate binding | Salmonella enterica subsp. enterica serovar Typhimurium |
| E214Q | increases KD value for quinolinic acid by 2fold and for 5-phospho-alpha-D-ribose 1-diphosphate by 2fold, has only modest effects on ligand binding and catalysis, wild-type-like pH profile. Presence of benzene-1,2-dicarboxylic acid results in 2fold tightening of 5-phospho-alpha-D-ribose 1-diphosphate binding | Salmonella enterica subsp. enterica serovar Typhimurium |
| K153A | inactive enzyme, kcat value decreases by more than 4000fold, is able to bind quinolinic acid with a KD value 2fold higher than that of wild-type | Salmonella enterica subsp. enterica serovar Typhimurium |
| K284A | KD value for quinolinic acid is similar to wild-type, decreases kcat by 30fold and increases Km and KD values for 5-phospho-alpha-D-ribose 1-diphosphate by 80fold and at least 20fold, respectively | Salmonella enterica subsp. enterica serovar Typhimurium |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.027 | - |
quinolinic acid | wild-type | Salmonella enterica subsp. enterica serovar Typhimurium |  |
| 0.041 | - |
quinolinic acid | mutant E214Q | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.057 | - |
5-phospho-alpha-D-ribose 1-diphosphate | wild-type | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.12 | - |
quinolinic acid | mutant E214D | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.121 | - |
quinolinic acid | mutant E214A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.128 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214Q | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.55 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214D | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.604 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 1.031 | - |
quinolinic acid | mutant D235A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 1.138 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant D235A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 1.463 | - |
quinolinic acid | mutant K284A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 4.12 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant K284A | Salmonella enterica subsp. enterica serovar Typhimurium | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Salmonella enterica subsp. enterica serovar Typhimurium | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| mutants purified to homogeneity in 250 mg yield from a 6 liter culture, by gel filtration | Salmonella enterica subsp. enterica serovar Typhimurium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| quinolinic acid + 5-phospho-alpha-D-ribose 1-diphosphate | Lys153 is necessary for catalysis and important for 5-phospho-alpha-D-ribose 1-diphosphate binding, Glu214 provides a hydrogen bond necessary for catalysis but does not act as a base or electrostatically to stabilize the transition state, Lys284 is involved in 5-phospho-alpha-D-ribose 1-diphosphate binding, and Asp235 is not essential | Salmonella enterica subsp. enterica serovar Typhimurium | nicotinic acid mononucleotide + diphosphate + CO2 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | wild-type and mutants, gel filtration | Salmonella enterica subsp. enterica serovar Typhimurium |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NadC | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| QAPRTase | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
| quinolinate phosphoribosyltransferase | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00024 | - |
quinolinic acid | mutant E214A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.00025 | - |
quinolinic acid | mutant E214D | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.033 | - |
quinolinic acid | mutant K284A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.5 | - |
quinolinic acid | mutant E214Q | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 1 | - |
quinolinic acid | wild-type | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 1.6 | - |
quinolinic acid | mutant D235A | Salmonella enterica subsp. enterica serovar Typhimurium | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00004 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.000049 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214D | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0007 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant K284A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0019 | - |
quinolinic acid | mutant E214A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.0019 | - |
quinolinic acid | mutant E214D | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 0.023 | - |
quinolinic acid | mutant K284A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 3.9 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant E214Q | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 12 | - |
5-phospho-alpha-D-ribose 1-diphosphate | mutant D235A | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 12 | - |
quinolinic acid | mutant E214Q | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 18 | - |
5-phospho-alpha-D-ribose 1-diphosphate | wild-type | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 37 | - |
quinolinic acid | wild-type | Salmonella enterica subsp. enterica serovar Typhimurium | |
| 52 | - |
quinolinic acid | mutant D235A | Salmonella enterica subsp. enterica serovar Typhimurium | |
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