Busca avançada
Ano de início
Entree
(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Redox Activation of Nox1 (NADPH Oxidase 1) Involves an Intermolecular Disulfide Bond Between Protein Disulfide Isomerase and p47(phox) in Vascular Smooth Muscle Cells

Texto completo
Autor(es):
Mostrar menos -
Gimenez, Marcela [1, 2] ; Verissimo-Filho, Sidney [1] ; Wittig, Ilka [3] ; Schickling, Brandon M. [2, 4] ; Hahner, Fabian [5] ; Schuermann, Christoph [5] ; Netto, Luis E. S. [6] ; Rosa, Jose Cesar [7, 8] ; Brandes, Ralf P. [5] ; Sartoretto, Simone [1, 4] ; Camargo, Livia De Lucca [1] ; Abdulkader, Fernando [9] ; Miller, Jr., Francis J. [2, 4, 10] ; Lopes, Lucia Rossetti [1]
Número total de Autores: 14
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Biomed Sci, Dept Pharmacol, Sao Paulo - Brazil
[2] Univ Iowa, Dept Med, Iowa City, IA 52242 - USA
[3] Goethe Univ, Funct Prote Core Unit, Frankfurt - Germany
[4] Duke Univ, Dept Med, Durham, NC - USA
[5] Goethe Univ, Inst Cardiovasc Physiol, Frankfurt - Germany
[6] Univ Sao Paulo, Dept Genet & Evolutionary Biol, Inst Biosci, Sao Paulo - Brazil
[7] Univ Sao Paulo, Dept Cell & Mol Biol, Ribeirao Preto Med Sch, Sao Paulo - Brazil
[8] Univ Sao Paulo, Pathogen Bioagents, Ribeirao Preto Med Sch, Sao Paulo - Brazil
[9] Univ Sao Paulo, Inst Biomed Sci, Dept Physiol & Biophys, Sao Paulo - Brazil
[10] Vet Affairs Med Ctr, Dept Med, Durham, NC - USA
Número total de Afiliações: 10
Tipo de documento: Artigo Científico
Fonte: ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY; v. 39, n. 2, p. 224-236, FEB 2019.
Citações Web of Science: 1
Resumo

Objective- PDI (protein disulfide isomerase A1) was reported to support Nox1 (NADPH oxidase) activation mediated by growth factors in vascular smooth muscle cells. Our aim was to investigate the molecular mechanism by which PDI activates Nox1 and the functional implications of PDI in Nox1 activation in vascular disease. Approach and Results- Using recombinant proteins, we identified a redox interaction between PDI and the cytosolic subunit p47(phox) in vitro. Mass spectrometry of crosslinked peptides confirmed redox-dependent disulfide bonds between cysteines of p47(phox) and PDI and an intramolecular bond between Cys 196 and 378 in p47(phox). PDI catalytic Cys 400 and p47(phox) Cys 196 were essential for the activation of Nox1 by PDI in vascular smooth muscle cells. Transfection of PDI resulted in the rapid oxidation of a redox-sensitive protein linked to p47(phox), whereas PDI mutant did not promote this effect. Mutation of p47(phox) Cys 196, or the redox active cysteines of PDI, prevented Nox1 complex assembly and vascular smooth muscle cell migration. Proximity ligation assay confirmed the interaction of PDI and p47(phox) in murine carotid arteries after wire injury. Moreover, in human atheroma plaques, a positive correlation between the expression of PDI and p47(phox) occurred only in PDI family members with the a ` redox active site. Conclusions- PDI redox cysteines facilitate Nox1 complex assembly, thus identifying a new mechanism through which PDI regulates Nox activity in vascular disease. (AU)

Processo FAPESP: 12/12033-8 - Estudo dos mecanismos moleculares envolvidos na interação dissulfeto isomerase proteica (PDI) e NADPH oxidase: papel da subunidade reguladora p47phox
Beneficiário:Marcela Gimenez
Linha de fomento: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 13/07937-8 - Redoxoma
Beneficiário:Ohara Augusto
Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
Processo FAPESP: 13/03520-5 - Papel da proteína dissulfeto isomerase na geração de espécies reativas de oxigênio pela NADPH oxidase durante o desenvolvimento da hipertensão arterial
Beneficiário:Lucia Rossetti Lopes
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 09/54764-6 - Regulação da homeostase redox e resposta integrada a estresse pela dissulfeto isomerase protéica (PDI): mecanismos e papel na fisiopatologia e terapêutica de doenças vasculares
Beneficiário:Francisco Rafael Martins Laurindo
Linha de fomento: Auxílio à Pesquisa - Temático