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Literature summary for 2.4.2.19 extracted from

  • Bello, Z.; Grubmeyer, C.
    Roles for cationic residues at the quinolinic acid binding site of quinolinate phosphoribosyltransferase (2010), Biochemistry, 49, 1388-1395.
    View publication on PubMedView publication on EuropePMC

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
K185A kcat value is reduced by 625fold, and binding affinity of quinolinic acid and 5-phospho-alpha-D-ribose 1-diphosphate to the enzyme decreases. Displays 83fold increase in activity toward the normally inactive quinolinic acid analogue, nicotinic acid. Displays a 300fold higher kcat/Km for nicotinic acid over the natural substrate quinolinic acid Salmonella enterica subsp. enterica serovar Typhimurium
additional information mutants can use 6-aminonicotinic acid as substrate, whereas 2,4-pyridinedicarboxylic acid, 2,5-pyridinedicarboxylic acid, 2,6-pyridinedicarboxylic acid, 3,4-pyridinedicarboxylic acid, 3,5-pyridinedicarboxylic acid, nicotinamide and pyridine-4-carboxylic acid are not utilized by the mutant enzymes Salmonella enterica subsp. enterica serovar Typhimurium
R118A results in 5000fold decrease in kcat value and a decrease in the binding affinity of quinolinic acid and 5-phospho-alpha-D-ribose 1-diphosphate to mutant R152A. Equimolar mixtures of mutant R118A with inactive or virtually inactive mutants produce approximately 50% of the enzymatic activity of wild-type Salmonella enterica subsp. enterica serovar Typhimurium
R152A kcat value is reduced by 33fold, and binding affinity of quinolinic acid and 5-phospho-alpha-D-ribose 1-diphosphate to the enzyme decreases. Displays 116fold increase in activity toward the normally inactive quinolinic acid analogue, nicotinic acid Salmonella enterica subsp. enterica serovar Typhimurium
R175A no activity 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, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.057
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.067
-
quinolinic acid mutant H188A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.136
-
quinolinic acid mutant R118A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.151
-
nicotinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.278
-
quinolinic acid mutant R152A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.547
-
5-phospho-alpha-D-ribose 1-diphosphate mutant H188A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.832
-
5-phospho-alpha-D-ribose 1-diphosphate mutant R152A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
1.478
-
quinolinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
2.181
-
5-phospho-alpha-D-ribose 1-diphosphate mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
3.138
-
5-phospho-alpha-D-ribose 1-diphosphate mutant R118A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium

Organism

Organism UniProt Comment Textmining
Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-

Purification (Commentary)

Purification (Comment) Organism
mutants purified by gel filtration, to homogeneity Salmonella enterica subsp. enterica serovar Typhimurium

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0006
-
wild-type with nicotinic acid at 1 mM and 2 mM 5-phospho-alpha-D-ribose 1-diphosphate Salmonella enterica subsp. enterica serovar Typhimurium
0.07
-
mutant R152A with nicotinic acid at 1 mM and 2 mM 5-phospho-alpha-D-ribose 1-diphosphate Salmonella enterica subsp. enterica serovar Typhimurium
0.1
-
mutant R152A with quinolinic acid at 1 mM and 2 mM 5-phospho-alpha-D-ribose 1-diphosphate Salmonella enterica subsp. enterica serovar Typhimurium
1.2
-
wild-type with quinolinic acid at 1 mM and 2 mM 5-phospho-alpha-D-ribose 1-diphosphate Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
Salmonella enterica subsp. enterica serovar Typhimurium beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
?
quinolinic acid + 5-phospho-alpha-D-ribose 1-diphosphate Arg175 is essential for catalysis, critical role for Lys185 in the chemistry of the QAPRTase intermediate, interfacial role for Arg118 during catalysis Salmonella enterica subsp. enterica serovar Typhimurium nicotinic acid mononucleotide + diphosphate + CO2
-
?

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 R118A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.0016
-
quinolinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.03
-
quinolinic acid mutant R152A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.048
-
nicotinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
1
-
quinolinic acid mutant H188A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
1
-
quinolinic acid wild-type, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium

General Information

General Information Comment Organism
physiological function overexpression and purification of the mutant enzymes produce 200-250 mg of protein/6 liter of culture similar to that of the wild-type enzyme 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
0.000064
-
5-phospho-alpha-D-ribose 1-diphosphate mutant R118A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.00073
-
5-phospho-alpha-D-ribose 1-diphosphate mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.0011
-
quinolinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.0015
-
quinolinic acid mutant R118A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.035
-
5-phospho-alpha-D-ribose 1-diphosphate mutant R152A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.1
-
quinolinic acid mutant R152A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
0.32
-
nicotinic acid mutant K185A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
1.8
-
5-phospho-alpha-D-ribose 1-diphosphate mutant H188A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
16
-
quinolinic acid mutant H188A, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
18
-
5-phospho-alpha-D-ribose 1-diphosphate wild-type, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium
37
-
quinolinic acid wild-type, at 25°C Salmonella enterica subsp. enterica serovar Typhimurium