Artículo
Biochemical characterization of GAF domain of free-R-methionine sulfoxide reductase from Trypanosoma cruzi
Gonzalez, Lihue Nadia
; Cabeza, Matías Sebastián
; Robello, Carlos; Guerrero, Sergio Adrian
; Iglesias, Alberto Alvaro
; Arias, Diego Gustavo
Fecha de publicación:
07/2023
Editorial:
Elsevier France-Editions Scientifiques Medicales Elsevier
Revista:
Biochimie
ISSN:
0300-9084
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Trypanosoma cruzi is the causal agent of Chagas Disease and is a unicellular parasite that infects a wide variety of mammalian hosts. The parasite exhibits auxotrophy by L-Met; consequently, it must be acquired from the extracellular environment of the host, either mammalian or invertebrate. Methionine (Met) oxidation produces a racemic mixture (R and S forms) of methionine sulfoxide (MetSO). Reduction of L-MetSO (free or protein-bound) to L-Met is catalyzed by methionine sulfoxide reductases (MSRs). Bioinformatics analyses identified the coding sequence for a free-R-MSR (fRMSR) enzyme in the genome of T. cruzi Dm28c. Structurally, this enzyme is a modular protein with a putative N-terminal GAF domain linked to a C-terminal TIP41 motif. We performed detailed biochemical and kinetic characterization of the GAF domain of fRMSR in combination with mutant versions of specific cysteine residues, namely, Cys12, Cys98, Cys108, and Cys132. The isolated recombinant GAF domain and full-length fRMSR exhibited specific catalytic activity for the reduction of free L-Met(R)SO (non-protein bound), using tryparedoxins as reducing partners. We demonstrated that this process involves two Cys residues, Cys98 and Cys132. Cys132 is the essential catalytic residue on which a sulfenic acid intermediate is formed. Cys98 is the resolutive Cys, which forms a disulfide bond with Cys132 as a catalytic step. Overall, our results provide new insights into redox metabolism in T. cruzi, contributing to previous knowledge of L-Met metabolism in this parasite.
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Articulos(CCT - SANTA FE)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Articulos(IAL)
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Citación
Gonzalez, Lihue Nadia; Cabeza, Matías Sebastián; Robello, Carlos; Guerrero, Sergio Adrian; Iglesias, Alberto Alvaro; et al.; Biochemical characterization of GAF domain of free-R-methionine sulfoxide reductase from Trypanosoma cruzi; Elsevier France-Editions Scientifiques Medicales Elsevier; Biochimie; 213; 7-2023; 190-204
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