Literature summary for 2.5.1.18 extracted from

Mathieu, Y.; Prosper, P.; Buee, M.; Dumarcay, S.; Favier, F.; Gelhaye, E.; Gerardin, P.; Harvengt, L.; Jacquot, J.P.; Lamant, T.; Meux, E.; Mathiot, S.; Didierjean, C.; Morel, M.
Characterization of a Phanerochaete chrysosporium glutathione transferase reveals a novel structural and functional class with ligandin properties (2012), J. Biol. Chem., 287, 39001-39011.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant GST5118 in Escherichia coli strain BL21(DE3), production of the selenomethionine-substituted GST5118 in a methionine auxotroph strain BL21(DE3)Met-	Phanerodontia chrysosporium

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant selenomethionine-apoGST5118 and native holoGST5118 with bound glutathione, microbatch under oil, paraffin, method at 4°C, 4 Selenomethionine-apoGST5118 crystals appear after 4-5 days from droplets containing 0.002 ml of 15-20 mg/ml protein in TE buffer, and 0.002 ml of precipitating solution containing 30% PEG 8000, 200 mM sodium acetate,and 100 mM sodium cacodylate, pH 6.5. The best crystals of GST5118 in complex with GSH grow after 2 months in droplets containing a mixture of 14 mg/ml protein and 6 mM GSH in TE buffer and precipitating solution containing 30% PEG 8000, 200 mM sodium acetate, and 100 mM HEPES, pH 7.0. X-ray diffraction structure determination and analysis at 1.8-2.0 A resolution, single-wavelength anomalous dispersion, the molecular replacement method fails to solve the protein structure	Phanerodontia chrysosporium

Crystallization (Comment)

Organism

purified recombinant selenomethionine-apoGST5118 and native holoGST5118 with bound glutathione, microbatch under oil, paraffin, method at 4°C, 4 Selenomethionine-apoGST5118 crystals appear after 4-5 days from droplets containing 0.002 ml of 15-20 mg/ml protein in TE buffer, and 0.002 ml of precipitating solution containing 30% PEG 8000, 200 mM sodium acetate,and 100 mM sodium cacodylate, pH 6.5. The best crystals of GST5118 in complex with GSH grow after 2 months in droplets containing a mixture of 14 mg/ml protein and 6 mM GSH in TE buffer and precipitating solution containing 30% PEG 8000, 200 mM sodium acetate, and 100 mM HEPES, pH 7.0. X-ray diffraction structure determination and analysis at 1.8-2.0 A resolution, single-wavelength anomalous dispersion, the molecular replacement method fails to solve the protein structure

Phanerodontia chrysosporium

Protein Variants

Protein Variants	Comment	Organism
S22A	site-directed mutagenesis	Phanerodontia chrysosporium
S22C	site-directed mutagenesis	Phanerodontia chrysosporium

Inhibitors

Inhibitors	Comment	Organism	Structure
Vanillin	-	Phanerodontia chrysosporium

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	GST5118 kinetic mechanism, overview	Phanerodontia chrysosporium
0.119	-	phenethyl isothiocyanate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.191	-	4-nitrophenyl butyrate	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.216	-	glutathione	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.615	-	glutathione	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.654	-	1-chloro-2,4-dinitrobenzene	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.774	-	4-nitrophenyl butyrate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium

Organism

Organism	UniProt	Comment	Textmining
Phanerodontia chrysosporium	I6ZUH3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant GST5118 from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, gel filtration, and anion exchange chromatography, followed by dialysis and ultrafiltration	Phanerodontia chrysosporium

Reaction

Reaction	Comment	Organism	Reaction ID
RX + glutathione = HX + R-S-glutathione	substrate binding structure and reaction mechanism, conformational changes, detailed overview	Phanerodontia chrysosporium	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
1-chloro-2,4-dinitrobenzene + glutathione	-	Phanerodontia chrysosporium	S-(2,4-dinitrophenyl)glutathione + HCl	-	?
4-nitrophenyl butyrate + glutathione	-	Phanerodontia chrysosporium	S-(4-nitrophenyl)glutathione + butyrate	-	?
additional information	GST5118 binds glutathionylated and sulfated compounds but also wood extractive molecules, such as vanillin, chloronitrobenzoic acid, hydroxyacetophenone, catechins, and aldehydes, in the glutathione pocket, overview. Glutathione binding structure and mechanism, detailed overview. The enzyme also shows peroxidase activity	Phanerodontia chrysosporium	?	-	?
phenethyl isothiocyanate + glutathione	-	Phanerodontia chrysosporium	?	-	?

Subunits

Subunits	Comment	Organism
homodimer	the GST5118 monomer adopts the GST canonical fold, which consists of an N-terminal thioredoxin-like motif and a C-terminal domain of at least four helices, unique dimeric organization of GST5118, overview	Phanerodontia chrysosporium

Synonyms

Synonyms	Comment	Organism
glutathione S-transferase	-	Phanerodontia chrysosporium
GST	-	Phanerodontia chrysosporium
GST5118	-	Phanerodontia chrysosporium

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.23	-	glutathione	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
0.29	-	4-nitrophenyl butyrate	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
8.52	-	phenethyl isothiocyanate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
11.03	-	glutathione	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
27.6	-	4-nitrophenyl butyrate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
29.42	-	1-chloro-2,4-dinitrobenzene	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	8	assay at	Phanerodontia chrysosporium

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.16	-	Vanillin	pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium

General Information

General Information	Comment	Organism
evolution	GST5118 belongs to a distinct class named GSTFuA, for fungal specific GST class A, as part of the GST superfamily	Phanerodontia chrysosporium
physiological function	glutathione S-transferases form a superfamily of multifunctional proteins with essential roles in cellular detoxification processes. GST5118 of Phanerochaete chrysosporium can function as a classical GST through the addition of glutathione mainly to phenethyl isothiocyanate, but alternatively and in a competitive way, it can also act as a ligandin of wood extractive compounds	Phanerodontia chrysosporium

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.37	-	glutathione	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
1.53	-	4-nitrophenyl butyrate	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
3.5	-	4-nitrophenyl butyrate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
44.83	-	1-chloro-2,4-dinitrobenzene	recombinant mutant S22A, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
51.67	-	glutathione	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium
71.67	-	phenethyl isothiocyanate	recombinant wild-type, pH 6.5, temperature not specified in the publication	Phanerodontia chrysosporium