Functional characterization of monothiol and dithiol glutaredoxins from Leptospira interrogans

Thiol redox proteins and low molecular mass thiols have essential functions in maintaining cellular redox balance in almost all living organisms. In the pathogenic bacterium Leptospira interrogans, several redox components have been described, namely, typical 2-Cys peroxiredoxin, a functional thiore...

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Bibliographic Details
Published in:Biochimie 2022-06, Vol.197, p.144-159
Main Authors: Sasoni, Natalia, Hartman, Matías D., García-Effron, Guillermo, Guerrero, Sergio A., Iglesias, Alberto A., Arias, Diego G.
Format: Article
Language:English
Subjects:
glutaredoxin
Glutathione
Iron-sulfur cluster
Leptospira
Redox metabolism
Thioredoxin
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Summary:Thiol redox proteins and low molecular mass thiols have essential functions in maintaining cellular redox balance in almost all living organisms. In the pathogenic bacterium Leptospira interrogans, several redox components have been described, namely, typical 2-Cys peroxiredoxin, a functional thioredoxin system, glutathione synthesis pathway, and methionine sulfoxide reductases. However, until now, information about proteins linked to GSH metabolism has not been reported in this pathogen. Glutaredoxins (Grxs) are GSH-dependent oxidoreductases that regulate and maintain the cellular redox state together with thioredoxins. This work deals with recombinant production at a high purity level, biochemical characterization, and detailed kinetic and structural study of the two Grxs (Lin1CGrx and Lin2CGrx) identified in L. interrogans serovar Copenhageni strain Fiocruz L1-130. Both recombinant LinGrxs exhibited the classical in vitro GSH-dependent 2-hydroxyethyl disulfide and dehydroascorbate reductase activity. Strikingly, we found that Lin2CGrx could serve as a substrate of methionine sulfoxide reductases A1 and B from L. interrogans. Distinctively, only recombinant Lin1CGrx contained a [2Fe2S] cluster confirming a homodimeric structure. The functionality of both LinGrxs was assessed by yeast complementation in null grx mutants, and both isoforms were able to rescue the mutant phenotype. Finally, our data suggest that protein glutathionylation as a post-translational modification process is present in L. interrogans. As a whole, our results support the occurrence of two new redox actors linked to GSH metabolism and iron homeostasis in L. interrogans. [Display omitted] •LinGrxs exhibited GSH-dependent thiol-disulfide reductase activity with HEDS.•Lin2CGrx is a reducing substrate of LinMsrB and LinMsrA1.•Lin1CGrx is a [Fe–S] cluster binding protein.•LinGrxs show cytoplasmic localization in L. interrogans.
ISSN:0300-9084
1638-6183
DOI:10.1016/j.biochi.2022.02.006