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Literature summary for 6.3.4.21 extracted from

  • Rajavel, M.; Lalo, D.; Gross, J.W.; Grubmeyer, C.
    Conversion of a cosubstrate to an inhibitor. Phosphorylation mutants of nicotinic acid phosphoribosyltransferase (1998), Biochemistry, 37, 4181-4188.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Saccharomyces cerevisiae

General Stability

General Stability Organism
first order activity loss in the presence of 1/200 trypsin/enzyme Salmonella enterica subsp. enterica serovar Typhimurium
sensitive to trypsin, partial proteolysis, 4fold increase in t1/2 for inactivation in the presence of 1 mM MgATP2- Salmonella enterica subsp. enterica serovar Typhimurium

Inhibitors

Inhibitors Comment Organism Structure
ATP
-
Saccharomyces cerevisiae
ATP 3 mM, competitive inhibition of nicotinic acid mononucleotide formation in mutants enzymes Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
nicotinate
-
Salmonella enterica subsp. enterica serovar Typhimurium
0.33 0.4 nicotinate mutant enzymes Salmonella enterica subsp. enterica serovar Typhimurium
2.3 48 5-phospho-alpha-D-ribose 1-diphosphate mutant enzymes Salmonella enterica subsp. enterica serovar Typhimurium
4.5
-
5-phospho-alpha-D-ribose 1-diphosphate
-
Salmonella enterica subsp. enterica serovar Typhimurium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45500
-
mutant enzyme, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium
45529
-
1 * 45529, gel filtration Salmonella enterica subsp. enterica serovar Typhimurium
45540
-
SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium
45550
-
mutant enzyme, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P39683
-
-
Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-

Storage Stability

Storage Stability Organism
4ΒΊC, as 70% saturated ammonium sulfate suspensions in 200 mM phosphate buffer, pH 8, 10% glycerol and 5 mM DTT Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information
-
Saccharomyces cerevisiae ?
-
?
additional information negligible ATPase activity in the absence of nicotinic acid Salmonella enterica subsp. enterica serovar Typhimurium ?
-
?
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O
-
Saccharomyces cerevisiae beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
?
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O the enzyme is a facultative ATPase that uses ATP hydrolysis to drive the synthesis of nicotinate mononucleotide and diphosphate from nicotinic acid and phosphoribosyl diphosphate in a steady-state reaction, the enzyme undergoes phosphorylation in His-219 by bound ATP, phosphorylated enzyme has higher affinity for substrates than does nonphosphorylated enzyme Salmonella enterica subsp. enterica serovar Typhimurium beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
?
nicotinate + 5-phospho-alpha-D-ribose 1-diphosphate + ATP + H2O mutants in the His-219 residue do catalyze the slow formation of nicotinic acid mononucleotide in the absence of ATP similarly to the wild type enzymes Salmonella enterica subsp. enterica serovar Typhimurium beta-nicotinate D-ribonucleotide + diphosphate + ADP + phosphate
-
?

Subunits

Subunits Comment Organism
monomer 1 * 45529, gel filtration Salmonella enterica subsp. enterica serovar Typhimurium

Cofactor

Cofactor Comment Organism Structure
ATP 1 mol ATP is hydrolyzed to ADP plus phosphate per mol product formed Salmonella enterica subsp. enterica serovar Typhimurium
ATP 1 mol ATP is hydrolyzed to ADP plus phosphate per mol product formed Saccharomyces cerevisiae
ATP the Vmax of nicotinic acid mononucleotide synthesis activity is stimulated 10fold Salmonella enterica subsp. enterica serovar Typhimurium