Human mitochondrial peroxiredoxin Prdx3 is dually localized in the intermembrane space and matrix subcompartments

Autor(es)

Gomes, Fernando ; Turano, Helena ; Haddad, Luciana A. ; Netto, Luis. E. S.

• Tipo de documento: Artigo Científico
• Fonte: REDOX BIOLOGY; v. 78, p. 13-pg., 2024-11-25.

Resumo

Peroxiredoxin 3 (Prdx3) is the major sink for H2O2 and other hydroperoxides within mitochondria, yet the mechanisms guiding the import of its cytosolic precursor into mitochondrial sub-compartments remain elusive. Prdx3 is synthesized in the cytosol as a precursor with an N-terminal cleavable presequence, which is frequently proposed to target the protein exclusively to the mitochondrial matrix. Here, we present a comprehensive analysis of the human Prdx3 biogenesis, using highly purified mitochondria from HEK293T cells. Sub- fractionation and probing for specific mitochondrial markers confirmed Prdx3 localization in the matrix, while unexpectedly revealed its presence in the mitochondrial intermembrane space (IMS). Both matrix and IMS isoforms were found to be soluble proteins, as demonstrated by alkaline carbonate extraction. By combining in silico analysis, in organello import assays and heterologous expression in yeast, we found that Prdx3 undergoes sequential proteolytic processing steps by mitochondrial processing peptidase (MPP) and mitochondrial intermediate peptidase (MIP) during its import into the matrix. Additionally, heterologous expression of Prdx3 in yeast revealed that its sorting to the IMS is dependent on the inner membrane peptidase (IMP) complex. Collectively, these findings uncover a complex submitochondrial distribution of Prdx3, supporting its multifaceted role in mitochondrial H2O2 metabolism. (AU)

• Processo FAPESP: 22/08446-7 - Mecanismos moleculares subjacentes à distribuição suborganelar de peroxirredoxinas mitocondriais de mamíferos: impactos na fisiologia e patologia
• Beneficiário: Luis Eduardo Soares Netto
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 23/01812-0 - Caracterização dos mecanismos moleculares envolvidos na importação mitocondrial da peroxirredoxina 3 (Prdx3) de mamíferos
• Beneficiário: Fernando Gomes
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico
• Processo FAPESP: 17/09443-3 - Importação de peroxirredoxinas para distintos compartimentos mitocondriais: impactos em fisiologia e patologia.
• Beneficiário: Fernando Gomes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/07937-8 - Redoxoma
• Beneficiário: Ohara Augusto
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 17/23839-7 - Investigação da ação bactericida da piocina S8: novas perspectivas para o tratamento de infecções causadas por cepas de Pseudomonas aeruginosa multirresistentes
• Beneficiário: Helena Gabriela Turano Gomes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado