Literature summary for 6.3.2.3 extracted from

Vergauwen, B.; De Vos, D.; Van Beeumen, J.J.
Characterization of the bifunctional gamma -glutamate-cysteine ligase-glutathione synthetase (GshF) of Pasteurella multocida (2005), J. Biol. Chem., 281, 4380-4394.

Search for more literature for 6.3.2.3

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli strain K12	Pasteurella multocida
expression in Escherichia coli	Pasteurella multocida

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.09	-	Gly	pH 8.2, 25°C	Pasteurella multocida
0.42	-	ATP	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida
0.53	-	ATP	pH 8.2, 25°C	Pasteurella multocida
5.9	-	gamma-Glu-L-Cys	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida
6.3	-	Gly	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida
81	-	gamma-Glu-L-Cys	pH 8.2, 25°C	Pasteurella multocida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pasteurella multocida
Mn2+	required	Pasteurella multocida

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
85000	-	2 * 85000, SDS-PAGE	Pasteurella multocida
85000	-	2 * 85000, gel filtration	Pasteurella multocida
85860	-	calculated from amino acid sequence	Pasteurella multocida
191000	-	bifunctional enzyme gamma-glutamate-cysteine ligase-glutathione synthetase, gel filtration	Pasteurella multocida
220000	-	about 220000 Da, gel filtration	Pasteurella multocida

Organism

Organism	UniProt	Comment	Textmining
Pasteurella multocida	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
Q-Sepharose FF HR 16/10 column chromatography, butyl-Sepharose FF HR 16/10 column chromatography, HiPrep 26/10 desalting column chromatography, Source Q anion-exchange resin chromatography, and Superdex G-200 (16/60) gel filtration	Pasteurella multocida
recombinant enzyme	Pasteurella multocida

Storage Stability

Storage Stability	Organism
-80°C, 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, can be stored for up to 6 months without significant loss of activity	Pasteurella multocida
-80°C, in 50 mM Tris-HCl (pH 7.4) containing100 mM NaCl, up to 6 months, without significant loss of activity	Pasteurella multocida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + gamma-Glu-L-Cys + Gly	-	Pasteurella multocida	ADP + phosphate + glutathione	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 85000, SDS-PAGE	Pasteurella multocida
dimer	2 * 85000, gel filtration	Pasteurella multocida

Synonyms

Synonyms	Comment	Organism
gamma -glutamate-cysteine ligase-glutathione synthetase	bifunctional enzyme	Pasteurella multocida
gamma-glutamate-cysteine ligase/glutathione synthetase	-	Pasteurella multocida
GshF	bifunctional enzyme	Pasteurella multocida
GshF	bifunctional enzyme gamma -glutamate-cysteine ligase-glutathione synthetase	Pasteurella multocida

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
25.5	-	ATP	pH 8.2, 25°C	Pasteurella multocida
26.9	-	gamma-Glu-L-Cys	pH 8.2, 25°C	Pasteurella multocida
36.3	-	Gly	pH 8.2, 25°C	Pasteurella multocida
55	-	ATP	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida
55	-	Gly	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida
55	-	gamma-Glu-L-Cys	inTris-HCl buffer (200 mM, pH 8.2)	Pasteurella multocida

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Pasteurella multocida

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Release 2023.1 (January 2023)