Literature summary for 2.5.1.18 extracted from

McManus, G.; Costa, M.; Canals, A.; Coll, M.; Mantle, T.J.
Site-directed mutagenesis of mouse glutathione transferase P1-1 unlocks masked cooperativity, introduces a novel mechanism for ping pong kinetic behaviour, and provides further structural evidence for participation of a water molecule in proton abstract (2011), FEBS J., 278, 273-281.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant GSTP1-1s in Escherichia coli	Mus musculus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant GSTP1-1 C47A mutant, hanging drop vapour diffusion method, mixing of 0.002 ml of 7 mg/ml protein in 10 mM MES, pH 6.0, with 0.0006 ml of 30 mM S-(p-nitrobenzyl)glutathione in 50 mM MES, pH 6.0, and 0.003 ml of reservoir solution containing 20% PEG 6000 and 0.1 M sodium citrate, pH 4.0, several weeks, 20°C, X-ray diffraction structure determmintion and analysis at 1.77 A resolution	Mus musculus

Crystallization (Comment)

Organism

purified recombinant GSTP1-1 C47A mutant, hanging drop vapour diffusion method, mixing of 0.002 ml of 7 mg/ml protein in 10 mM MES, pH 6.0, with 0.0006 ml of 30 mM S-(p-nitrobenzyl)glutathione in 50 mM MES, pH 6.0, and 0.003 ml of reservoir solution containing 20% PEG 6000 and 0.1 M sodium citrate, pH 4.0, several weeks, 20°C, X-ray diffraction structure determmintion and analysis at 1.77 A resolution

Mus musculus

Protein Variants

Protein Variants	Comment	Organism
C14A	site-directed mutagenesis	Mus musculus
C14A/C169A	site-directed mutagenesis	Mus musculus
C14A/C47A	site-directed mutagenesis	Mus musculus
C169A	site-directed mutagenesis	Mus musculus
C47A	site-directed mutagenesis	Mus musculus
C47A/C169A	site-directed mutagenesis	Mus musculus
additional information	the native enzyme and the C169A mutant exhibit Michaelis-Menten kinetics, but all other thiol to alanine mutants exhibit sigmoidal kinetics to varying degrees	Mus musculus

Inhibitors

Inhibitors	Comment	Organism	Structure
p-nitrobenzylglutathione	-	Mus musculus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	the native enzyme and the C169A mutant exhibit MichaelisMenten kinetics, but all other thiol to alanine mutants exhibit sigmoidal kinetics to varying degrees, ping-pong kinetics of mutant C169A, overview	Mus musculus

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	P19157	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant GSTP1-1s from Escherichia coli by glutathione affinity chromatography	Mus musculus

Reaction

Reaction	Comment	Organism	Reaction ID
RX + glutathione = HX + R-S-glutathione	the mutant C169A exhibits cooperative behaviour and a ping pong mechanism whereby liberation of a proton from a reduced enzyme-glutathione complex to form an enzyme-unprotonated glutathione complex is essentially irreversible	Mus musculus	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	-	Mus musculus	-

Synonyms

Synonyms	Comment	Organism
glutathione transferase P1-1	-	Mus musculus
GSTP1-1	-	Mus musculus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	-	assay at	Mus musculus