title: The glutaredoxin mono- and di-thiol mechanisms for deglutathionylation are functionally equivalent: implications for redox systems biology.
abstract: Glutathionylation plays a central role in cellular redox regulation and anti-oxidative defence. Grx (Glutaredoxins) are primarily responsible for reversing glutathionylation and their activity therefore affects a range of cellular processes, making them prime candidates for computational systems biology studies. However, two distinct kinetic mechanisms involving either one (monothiol) or both (dithiol) active-site cysteines have been proposed for their deglutathionylation activity and initial studies predicted that computational models based on either of these mechanisms will have different structural and kinetic properties. Further, a number of other discrepancies including the relative activity of active-site mutants and contrasting reciprocal plot kinetics have also been reported for these redoxins. Using kinetic modelling, we show that the dithiol and monothiol mechanisms are identical and, we were also able to explain much of the discrepant data found within the literature on Grx activity and kinetics. Moreover, our results have revealed how an apparently futile side-reaction in the monothiol mechanism may play a significant role in regulating Grx activity in vivo.
authors.affiliation: *School of Life Sciences, University of KwaZulu-Natal, Carbis Road, Pietermaritzburg, 3201, South Africa.
authors.forename: Lefentse N
authors.initials: LN
authors.lastname: Mashamaite
authors.affiliation: †Department of Biochemistry, Stellenbosch University, Private Bag X1, Matieland, 7602 Stellenbosch, South Africa.
authors.forename: Johann M
authors.initials: JM
authors.lastname: Rohwer
authors.affiliation: *School of Life Sciences, University of KwaZulu-Natal, Carbis Road, Pietermaritzburg, 3201, South Africa.
authors.forename: Ché S
authors.initials: CS
authors.lastname: Pillay
mesh_terms: D054477:Glutaredoxins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D049490:Systems Biology
chemical_list: D054477:Glutaredoxins; D029701:Saccharomyces cerevisiae Proteins; D013438:Sulfhydryl Compounds