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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Nitroarachidonic acid (NO(2)AA) inhibits protein disulfide isomerase (PDI) through reversible covalent adduct formation with critical cysteines

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Gonzalez-Perilli, Lucia ; Mastrogiovanni, Mauricio ; Fernandes, Denise de Castro ; Rubbo, Homero ; Laurindo, Francisco ; Trostchansky, Andres
Total Authors: 6
Document type: Journal article
Source: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1861, n. 5, A, p. 1131-1139, MAY 2017.
Web of Science Citations: 6

Background: Nitroarachidonic acid (NO(2)AA) exhibits pleiotropic anti-inflammatory actions in a variety of cell types. We have recently shown that NO(2)AA inhibits phagocytic NADPH oxidase 2 (NOX2) by preventing the formation of the active complex. Recent work indicates the participation of protein disulfide isomerase (PDI) activity in NOX2 activation. Cysteine (Cys) residues at PDI active sites could be targets for NO(2)AA- nitroalkylation regulating PDI activity which could explain our previous observation. Methods: PDI reductase and chaperone activities were assessed using the insulin and GFP renaturation methods in the presence or absence of NO(2)AA. To determine the covalent reaction with PDI as well as the site of reaction, the PEG-switch assay and LC-MS/MS studies were performed. Results and conclusions: We determined that both activities of PDI were inhibited by NO(2)AA in a dose- and time-dependent manner and independent from release of nitric oxide. Since nitroalkenes are potent electrophiles and PDI has critical Cys residues for its activity, then formation of a covalent adduct between NO(2)AA and PDI is feasible. To this end we demonstrated the reversible covalent modification of PDI by NO(2)AA. Trypsinization of modified PDI confirmed that the Cys residues present in the active site a' of PDI were key targets accounting for nitroalkene modification. General significance: PDI may contribute to NOX2 activation. As such, inhibition of PDI by NO(2)AA might be involved in preventing NOX2 activation. Future work will be directed to determine if the covalent modifications observed play a role in the reported NO(2)AA inhibition of NOX2 activity. (C) 2017 Elsevier B.V. All rights reserved. (AU)

FAPESP's process: 13/07937-8 - Redoxome - Redox Processes in Biomedicine
Grantee:Ohara Augusto
Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC