Literature summary for 2.4.2.10 extracted from

Gonzalez-Segura, L.; Witte, J.F.; McClard, R.W.; Hurley, T.D.
Ternary complex formation and induced asymmetry in orotate phosphoribosyltransferase (2007), Biochemistry, 46, 14075-14086.

Cloned(Commentary)

Cloned (Comment)	Organism
mutants are expressed in Escherichia coli strain CS101-4UI	Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment)	Organism
sitting drop vapour diffusion method, the apoenzyme is crystallized in 100 mM Tris-HCl, pH 7.4, 0.2 M magnesium acetate, and 26% (w/v) PEG 6000, crystals of OPRTase in complex with substrates (5.0 mM 5-phospho-alpha-D-ribose 1-diphosphate and 5.0 mM orotate) or product (5.0 mM orotidine 5'-phosphate) are grown in 100 mM sodium acetate, pH 4.6, 0.14 M ammonium acetate, and 38% (w/v) PEG 4000, crystals of the OMP complex are grown from solutions containing 100 mM Tris HCl, pH 7.4, 0.08 M magnesium acetate, and 28% (w/v) PEG 6000	Saccharomyces cerevisiae

Crystallization (Comment)

Organism

sitting drop vapour diffusion method, the apoenzyme is crystallized in 100 mM Tris-HCl, pH 7.4, 0.2 M magnesium acetate, and 26% (w/v) PEG 6000, crystals of OPRTase in complex with substrates (5.0 mM 5-phospho-alpha-D-ribose 1-diphosphate and 5.0 mM orotate) or product (5.0 mM orotidine 5'-phosphate) are grown in 100 mM sodium acetate, pH 4.6, 0.14 M ammonium acetate, and 38% (w/v) PEG 4000, crystals of the OMP complex are grown from solutions containing 100 mM Tris HCl, pH 7.4, 0.08 M magnesium acetate, and 28% (w/v) PEG 6000

Saccharomyces cerevisiae

Protein Variants

Protein Variants	Comment	Organism
D131A	severly reduced activity	Saccharomyces cerevisiae
D131A/D132A	severly reduced activity	Saccharomyces cerevisiae
D132A	severly reduced activity	Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.004	-	orotidine 5'-phosphate	wild type enzyme	Saccharomyces cerevisiae
0.007	-	orotidine 5'-phosphate	mutant enzyme D131A	Saccharomyces cerevisiae
0.007	-	orotidine 5'-phosphate	mutant enzyme D132A	Saccharomyces cerevisiae
0.01	-	orotidine 5'-phosphate	mutant enzyme D131A/D132A	Saccharomyces cerevisiae
0.041	-	5-phospho-alpha-D-ribose 1-diphosphate	wild type enzyme	Saccharomyces cerevisiae
0.043	-	5-phospho-alpha-D-ribose 1-diphosphate	mutant enzyme D131A	Saccharomyces cerevisiae
0.086	-	5-phospho-alpha-D-ribose 1-diphosphate	mutant enzyme D131A/D132A	Saccharomyces cerevisiae
0.12	-	5-phospho-alpha-D-ribose 1-diphosphate	mutant enzyme D132A	Saccharomyces cerevisiae

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	Mg2+-dependent activity	Saccharomyces cerevisiae

Organism

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	P13298	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
3	8	wild type enzyme	Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
orotate + 5-phospho-alpha-D-ribose 1-diphosphate	-	Saccharomyces cerevisiae	orotidine 5'-phosphate + diphosphate	-	?

Subunits

Subunits	Comment	Organism
homodimer	x-ray crystallography	Saccharomyces cerevisiae

Synonyms

Synonyms	Comment	Organism
OPRTase	-	Saccharomyces cerevisiae