Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins

The goal of the present study was to determine whether peptidylarginine deiminase PAD2 and PAD4 enzymes are present in Balb/c mouse salivary glands and whether they are able to citrullinate Ro and La ribonucleoproteins. Salivary glands from Balb/c mice were cultured in DMEM and supplemented with one...

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Main Authors: Mayra Rodríguez-Rodríguez, Rafael Herrera-Esparza, Juan-José Bollain y Goytia, María-Elena Pérez-Pérez, Deyanira Pacheco-Tovar, Jessica Murillo-Vázquez, Guadalupe Pacheco-Tovar, Esperanza Avalos-Díaz
Format: Article
Language:English
Published: Wiley 2017-01-01
Series:Journal of Immunology Research
Online Access:http://dx.doi.org/10.1155/2017/8959687
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author Mayra Rodríguez-Rodríguez
Rafael Herrera-Esparza
Juan-José Bollain y Goytia
María-Elena Pérez-Pérez
Deyanira Pacheco-Tovar
Jessica Murillo-Vázquez
Guadalupe Pacheco-Tovar
Esperanza Avalos-Díaz
author_facet Mayra Rodríguez-Rodríguez
Rafael Herrera-Esparza
Juan-José Bollain y Goytia
María-Elena Pérez-Pérez
Deyanira Pacheco-Tovar
Jessica Murillo-Vázquez
Guadalupe Pacheco-Tovar
Esperanza Avalos-Díaz
author_sort Mayra Rodríguez-Rodríguez
collection DOAJ
description The goal of the present study was to determine whether peptidylarginine deiminase PAD2 and PAD4 enzymes are present in Balb/c mouse salivary glands and whether they are able to citrullinate Ro and La ribonucleoproteins. Salivary glands from Balb/c mice were cultured in DMEM and supplemented with one of the following stimulants: ATP, LPS, TNF, IFNγ, or IL-6. A control group without stimulant was also evaluated. PAD2, PAD4, citrullinated peptides, Ro60, and La were detected by immunohistochemistry and double immunofluorescence. PAD2 and PAD4 mRNAs and protein expression were detected by qPCR and Western blot analysis. PAD activity was assessed using an antigen capture enzyme-linked immunosorbent assay. LPS, ATP, and TNF triggered PAD2 and PAD4 expression; in contrast, no expression was detected in the control group (p<0.001). PAD transcription slightly increased in response to stimulation. Additionally, PAD2/4 activity modified the arginine residues of a reporter protein (fibrinogen) in vitro. PADs citrullinated Ro60 and La ribonucleoproteins in vivo. Molecular stimulants induced apoptosis in ductal cells and the externalization of Ro60 and La ribonucleoproteins onto apoptotic membranes. PAD enzymes citrullinate Ro and La ribonucleoproteins, and this experimental approach may facilitate our understanding of the role of posttranslational modifications in the pathophysiology of Sjögren’s syndrome.
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spelling doaj-art-97db971ca90e47ea9a833475b200f2a82025-08-20T03:37:08ZengWileyJournal of Immunology Research2314-88612314-71562017-01-01201710.1155/2017/89596878959687Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La RibonucleoproteinsMayra Rodríguez-Rodríguez0Rafael Herrera-Esparza1Juan-José Bollain y Goytia2María-Elena Pérez-Pérez3Deyanira Pacheco-Tovar4Jessica Murillo-Vázquez5Guadalupe Pacheco-Tovar6Esperanza Avalos-Díaz7Department of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoPharmacology PhD Program, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Guadalajara, JAL, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoDepartment of Immunology, Unidad Académica de Ciencias Biológicas, Universidad Autónoma de Zacatecas, Guadalupe, ZAC, MexicoThe goal of the present study was to determine whether peptidylarginine deiminase PAD2 and PAD4 enzymes are present in Balb/c mouse salivary glands and whether they are able to citrullinate Ro and La ribonucleoproteins. Salivary glands from Balb/c mice were cultured in DMEM and supplemented with one of the following stimulants: ATP, LPS, TNF, IFNγ, or IL-6. A control group without stimulant was also evaluated. PAD2, PAD4, citrullinated peptides, Ro60, and La were detected by immunohistochemistry and double immunofluorescence. PAD2 and PAD4 mRNAs and protein expression were detected by qPCR and Western blot analysis. PAD activity was assessed using an antigen capture enzyme-linked immunosorbent assay. LPS, ATP, and TNF triggered PAD2 and PAD4 expression; in contrast, no expression was detected in the control group (p<0.001). PAD transcription slightly increased in response to stimulation. Additionally, PAD2/4 activity modified the arginine residues of a reporter protein (fibrinogen) in vitro. PADs citrullinated Ro60 and La ribonucleoproteins in vivo. Molecular stimulants induced apoptosis in ductal cells and the externalization of Ro60 and La ribonucleoproteins onto apoptotic membranes. PAD enzymes citrullinate Ro and La ribonucleoproteins, and this experimental approach may facilitate our understanding of the role of posttranslational modifications in the pathophysiology of Sjögren’s syndrome.http://dx.doi.org/10.1155/2017/8959687
spellingShingle Mayra Rodríguez-Rodríguez
Rafael Herrera-Esparza
Juan-José Bollain y Goytia
María-Elena Pérez-Pérez
Deyanira Pacheco-Tovar
Jessica Murillo-Vázquez
Guadalupe Pacheco-Tovar
Esperanza Avalos-Díaz
Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
Journal of Immunology Research
title Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
title_full Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
title_fullStr Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
title_full_unstemmed Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
title_short Activation of Peptidylarginine Deiminase in the Salivary Glands of Balb/c Mice Drives the Citrullination of Ro and La Ribonucleoproteins
title_sort activation of peptidylarginine deiminase in the salivary glands of balb c mice drives the citrullination of ro and la ribonucleoproteins
url http://dx.doi.org/10.1155/2017/8959687
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