title: Crystal Structure of Saccharomyces cerevisiae ECM4, a Xi-Class Glutathione Transferase that Reacts with Glutathionyl-(hydro)quinones.
abstract: Glutathionyl-hydroquinone reductases (GHRs) belong to the recently characterized Xi-class of glutathione transferases (GSTXs) according to unique structural properties and are present in all but animal kingdoms. The GHR ScECM4 from the yeast Saccharomyces cerevisiae has been studied since 1997 when it was found to be potentially involved in cell-wall biosynthesis. Up to now and in spite of biological studies made on this enzyme, its physiological role remains challenging. The work here reports its crystallographic study. In addition to exhibiting the general GSTX structural features, ScECM4 shows extensions including a huge loop which contributes to the quaternary assembly. These structural extensions are probably specific to Saccharomycetaceae. Soaking of ScECM4 crystals with GS-menadione results in a structure where glutathione forms a mixed disulfide bond with the cysteine 46. Solution studies confirm that ScECM4 has reductase activity for GS-menadione in presence of glutathione. Moreover, the high resolution structures allowed us to propose new roles of conserved residues of the active site to assist the cysteine 46 during the catalytic act.
authors.affiliation: Université de Lorraine, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.|CNRS, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Schwartz
authors.affiliation: Université de Lorraine, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.|CNRS, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Didierjean
authors.affiliation: Université de Lorraine, Interactions Arbres-Microorganismes, UMR1136, F-54500 Vandœuvre-lès-Nancy, France.|INRA, Interactions Arbres-Microorganismes, UMR1136, F-54280 Champenoux, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Hecker
authors.affiliation: Université de Lorraine, Interactions Arbres-Microorganismes, UMR1136, F-54500 Vandœuvre-lès-Nancy, France.|INRA, Interactions Arbres-Microorganismes, UMR1136, F-54280 Champenoux, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Girardet
authors.affiliation: Université de Lorraine, Interactions Arbres-Microorganismes, UMR1136, F-54500 Vandœuvre-lès-Nancy, France.|INRA, Interactions Arbres-Microorganismes, UMR1136, F-54280 Champenoux, France.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Morel-Rouhier
authors.affiliation: Université de Lorraine, Interactions Arbres-Microorganismes, UMR1136, F-54500 Vandœuvre-lès-Nancy, France.|INRA, Interactions Arbres-Microorganismes, UMR1136, F-54280 Champenoux, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Gelhaye
authors.affiliation: Université de Lorraine, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.|CNRS, CRM2, UMR 7036, F-54500 Vandœuvre-lès-Nancy, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Favier
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D055162:Biocatalysis; D018360:Crystallography, X-Ray; D005978:Glutathione / Q000737:chemistry; D005982:Glutathione Transferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011809:Quinones / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D024483:Vitamin K 3 / Q000737:chemistry / Q000378:metabolism
chemical_list: D011809:Quinones; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D024483:Vitamin K 3; D005982:Glutathione Transferase; D005978:Glutathione