Journal article
Redox Activation of Nox1 (NADPH Oxidase 1) Involves an Intermolecular Disulfide Bond Between Protein Disulfide Isomerase and p47phox in Vascular Smooth Muscle Cells
Arteriosclerosis, thrombosis, and vascular biology, Vol.39(2), pp.224-236
02/01/2019
DOI: 10.1161/ATVBAHA.118.311038
PMCID: PMC6344286
PMID: 30580571
Abstract
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.
Details
- Title: Subtitle
- Redox Activation of Nox1 (NADPH Oxidase 1) Involves an Intermolecular Disulfide Bond Between Protein Disulfide Isomerase and p47phox in Vascular Smooth Muscle Cells
- Creators
- Marcela Gimenez - University of IowaSidney Verissimo-Filho - Universidade de São PauloIlka Wittig - Goethe University FrankfurtBrandon M. Schickling - University of IowaFabian Hahner - Goethe University FrankfurtChristoph Schuermann - Goethe Univ, Inst Cardiovasc Physiol, Frankfurt, GermanyLuis E. S. Netto - Universidade de São PauloJose Cesar Rosa - Universidade de São PauloRalf P. Brandes - Goethe University FrankfurtSimone Sartoretto - Universidade de São PauloLivia De Lucca Camargo - Universidade de São PauloFernando Abdulkader - Universidade de São PauloFrancis J. Miller - University of IowaLucia Rossetti Lopes - Universidade de São Paulo
- Resource Type
- Journal article
- Publication Details
- Arteriosclerosis, thrombosis, and vascular biology, Vol.39(2), pp.224-236
- DOI
- 10.1161/ATVBAHA.118.311038
- PMID
- 30580571
- PMCID
- PMC6344286
- NLM abbreviation
- Arterioscler Thromb Vasc Biol
- ISSN
- 1079-5642
- eISSN
- 1524-4636
- Publisher
- Lippincott Williams & Wilkins
- Number of pages
- 13
- Grant note
- 2I01BX001729 / Office of Research and Development, Department of Veterans Affairs; US Department of Veterans Affairs 475609/2013-1 / Conselho Nacional de Desenvolvimento Cientifico Tecnologico (CNPq); Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPQ) 2012/12033-8 / FAPESP; Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) 2012/12033-8; 2013/183000; 2013/07937-8; 2009/54764-6; 2013/03520-5 / Fundacao de Amparo a Pesquisa do Estado de Sao Paulo; Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) HL130039 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA SFB815 / DFG; German Research Foundation (DFG) PRE29980013 / American Heart Association
- Language
- English
- Date published
- 02/01/2019
- Academic Unit
- Obstetrics and Gynecology
- Record Identifier
- 9984966747102771
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