title: Glyoxalase II promotes "in vitro" S-glutathionylation.
abstract: S-glutathionylation involves the reversible formation of a mix disulphide-bridge between specific cysteine and a molecule of glutathione, the major non-protein antioxidant compound in the cell. Mechanisms of protein S-glutathionylation are far to be completely understood and several reactions can promote it, either spontaneously or catalyzed. For the first time Glo II enzyme was studied as a new potential candidate to promote S-glutathionylation. To demonstrate its active involvement in protein glutathionylation were used actin, malate dehydrogenase and GAPDH purified proteins, which are known to be glutathionylated, for in vitro experiments..This work shows active involvement of cytosolic Glo II for in vitro protein S-glutathionylation. To confirm the role of Glo II, preliminary protein-protein docking studies was performed between Glo II and human actin. The data showed a high propensity to aggregate with other proteins through its catalytic site Further, in silico investigation of Glo II stability and behavior, conducted through full atom molecular dynamics simulations, showed an high folding stability together with a great affinity towards its own reaction product glutathione both protonated (GSH) and unprotonated (GS(-)). These studies, revealed that GloII, using its natural substrate SLG, allow a rapid and specific protein-SSG formation, leading enzymatic regulation of S-glutathionylation in proteins of different origin and cellular compartmentalization.
authors.affiliation: Polytechnic University of Marche (Ancona ), Department of Clinical Sciences, Section of Biochemistry, Biology and Physic, Italy. Electronic address: l.cianfruglia@univpm.it.
authors.forename: Laura
authors.initials: L
authors.lastname: Cianfruglia
authors.affiliation: Polytechinc University of Marche (Ancona ), Department of Life and Environmental Sciences, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Galeazzi
authors.affiliation: Polytechinc University of Marche (Ancona ), Department of Life and Environmental Sciences, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Massaccesi
authors.affiliation: Polytechnic University of Marche (Ancona ), Department of Clinical Sciences, Section of Biochemistry, Biology and Physic, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Spaccini
authors.affiliation: Polytechnic University of Marche (Ancona ), Department of Clinical Sciences, Section of Biochemistry, Biology and Physic, Italy.
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Caniglia
authors.affiliation: Polytechnic University of Marche (Ancona ), Department of Clinical Sciences, Section of Biochemistry, Biology and Physic, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Principato
authors.affiliation: Polytechnic University of Marche (Ancona ), Department of Clinical Sciences, Section of Biochemistry, Biology and Physic, Italy.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Armeni