Identification of glutathione (GSH)-independent glyoxalase III from Schizosaccharomyces pombe

Abstract

BackgroundReactive carbonyl species (RCS), such as methylglyoxal (MG) and glyoxal (GO), are synthesized as toxic metabolites in living systems. Mechanisms of RCS detoxification include the glutathione (GSH)-dependent system consisting of glyoxalase I (GLO1) and glyoxalase II (GLO2), and GSH-independent system involving glyoxalase III (GLO3). Hsp31 and DJ-1 proteins are weakly homologous to each other and belong to two different subfamilies of the DJ-1/Hsp31/PfpI superfamily. Recently, the Escherichia coli Hsp31 protein and the DJ-1 proteins from Arabidopsis thaliana and metazoans have been demonstrated to have GLO3 activity.ResultsWe performed a systematic survey of homologs of DJ-1 and Hsp31 in fungi. We found that DJ-1 proteins have a very limited distribution in fungi, whereas Hsp31 proteins are widely distributed among different fungal groups. Phylogenetic analysis revealed that fungal and metazoan DJ-1 proteins and bacterial YajL proteins are most closely related and together form a sister clade to bacterial and fungal Hsp31 proteins. We showed that two Schizosaccharomyces pombe Hsp31 proteins (Hsp3101 and Hsp3102) and one Saccharomyces cerevisiae Hsp31 protein (ScHsp31) displayed significantly higher in vitro GLO3 activity than S. pombe DJ-1 (SpDJ-1). Overexpression of hsp3101, hsp3102 and ScHSP31 could confer MG and GO resistance on either wild-type S. pombe cells or GLO1 deletion of S. pombe. S. pombe DJ-1 and Hsp31 proteins exhibit different patterns of subcellular localization.ConclusionsOur results suggest that fungal Hsp31 proteins are the major GLO3 that may have some role in protecting cells from RCS toxicity in fungi. Our results also support the view that the GLO3 activity of Hsp31 proteins may have evolved independently from that of DJ-1 proteins.

Robust Glyoxalase activity of Hsp31, a ThiJ/DJ-1/PfpI Family Member Protein, Is Critical for Oxidative Stress Resistance in Saccharomyces cerevisiae

“…2A). This observation was consistent with the previous observations in Hsp3101 of Schizosaccharomyces pombe and Glx3 of Candida albicans (58,59). The protein exhibited typical Michaelis-Menten enzyme kinetics with an estimated K m of 3.854 ϫ 10 Ϫ4 M and V max of 3.073 ϫ 10 Ϫ2 M. Interestingly, the catalytic constant (K cat ) of yeast Hsp31 was found to be 150 min Ϫ1 , which FIGURE 1.…”

Epidermal Growth Factor-dependent Activation of the Extracellular Signal-regulated Kinase Pathway by DJ-1 Protein through Its Direct Binding to c-Raf Protein

“…6), indicating that DJ-1 is able to activate c-Raf without other factors under these conditions. Although DJ-1 has several enzymatic activities, including protease (37)(38)(39)(40) and glyoxalase activities (41)(42)(43)(44), there is no report showing that DJ-1 is a protein kinase and that DJ-1 has a structural motif for kinase (3)(4)(5). These results therefore suggest that DJ-1 enhances self-phosphorylation activity of c-Raf without a role as kinase.…”

Concomitant Loss of the Glyoxalase System and Glycolysis Makes the Uncultured Pathogen “Candidatus Liberibacter asiaticus” an Energy Scavenger

“…In the case of B. burgdorferi, AKR is the sole predicted MG detoxification enzyme. GloIII, which can catalyze direct detoxification of MG to D-lactic acid (19)(20)(21), was omitted from Table 1 because GloIII (and similar proteins with a DJ-1 domain) are reportedly present at relatively low cellular concentrations and have extremely low specific activity, thereby precluding GloIII activity as a significant MG detoxification mechanism in vivo (23). Based on TBLASTN and keyword searches, all pathogenic and uncultured liberibacters sequenced to date appear to lack all previously described MG detoxification pathways.…”

“…Potential alternative MG detoxification enzymes have been described. For example, a metal ion-and glutathione (GSH)-independent route for direct detoxification of MG to D-lactic acid is catalyzed by glyoxalase III (GloIII; EC 4.2.1.130) in Escherichia coli (19), Schizosaccharomyces pombe (20), and rice (21). Nonglyoxalase enzymes such as aldehyde reductase (AR; EC 1.1.1.2), aldehyde dehydrogenase (AD; EC 1.2.1.3), and aldo/keto reductases (AKR superfamily; EC 1.1) can also potentially detoxify MG (22).…”