Disulfide stress: a novel type of oxidative stress in acute pancreatitis.
Glutathione oxidation and protein glutathionylation are considered hallmarks of oxidative stress in cells because they reflect thiol redox status in proteins. Our aims were to analyze the redox status of thiols and to identify mixed disulfides and targets of redox signaling in pancreas in experimental acute pancreatitis as a model of acute inflammation associated with glutathione depletion. Glutathione depletion in pancreas in acute pancreatitis is not associated with any increase in oxidized glutathione levels or protein glutathionylation. Cystine and homocystine levels as well as protein cysteinylation and γ-glutamyl cysteinylation markedly rose in pancreas after induction of pancreatitis. Protein cysteinylation was undetectable in pancreas under basal conditions. Targets of disulfide stress were identified by Western blotting, diagonal electrophoresis, and proteomic methods. Cysteinylated albumin was detected. Redox-sensitive PP2A and tyrosine protein phosphatase activities diminished in pancreatitis and this loss was abrogated by N-acetylcysteine. According to our findings, disulfide stress may be considered a specific type of oxidative stress in acute inflammation associated with protein cysteinylation and γ-glutamylcysteinylation and oxidation of the pair cysteine/cystine, but without glutathione oxidation or changes in protein glutathionylation. Two types of targets of disulfide stress were identified: redox buffers, such as ribonuclease inhibitor or albumin, and redox-signaling thiols, which include thioredoxin 1, APE1/Ref1, Keap1, tyrosine and serine/threonine phosphatases, and protein disulfide isomerase. These targets exhibit great relevance in DNA repair, cell proliferation, apoptosis, endoplasmic reticulum stress, and inflammatory response. Disulfide stress would be a specific mechanism of redox signaling independent of glutathione redox status involved in inflammation.
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain.
Mari-Luz
ML
Moreno
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain; Division of Neonatology, University Hospital Materno-Infantil La Fe, 46026 Valencia, Spain.
Javier
J
Escobar
Servicio de Inmunología, Hospital Universitario de La Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid, Spain.
Alicia
A
Izquierdo-Álvarez
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain.
Anabel
A
Gil
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain.
Salvador
S
Pérez
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain.
Javier
J
Pereda
Servicio de Inmunología, Hospital Universitario de La Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid, Spain; Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de Madrid, Madrid, Spain.
Inés
I
Zapico
Division of Neonatology, University Hospital Materno-Infantil La Fe, 46026 Valencia, Spain.
Máximo
M
Vento
Department of Surgery, University Clinic Hospital, University of Valencia, 46010 Valencia, Spain.
Luis
L
Sabater
Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de Madrid, Madrid, Spain.
Anabel
A
Marina
Servicio de Inmunología, Hospital Universitario de La Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid, Spain.
Antonio
A
Martínez-Ruiz
Department of Physiology, School of Pharmacy, University of Valencia, 46100 Burjasot (Valencia), Spain. Electronic address: juan.sastre@uv.es.
Juan
J
Sastre
D000818:Animals; D003545:Cysteine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D005609:Free Radicals / Q000378:metabolism; D019803:Glutathione Disulfide / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010195:Pancreatitis / Q000378:metabolism* / Q000473:pathology; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D017510:Protein Folding; D013312:Stress, Physiological; D013438:Sulfhydryl Compounds / Q000378:metabolism
D004220:Disulfides; D005609:Free Radicals; D013438:Sulfhydryl Compounds; D019704:Protein Disulfide-Isomerases; D003545:Cysteine; D019803:Glutathione Disulfide
Cysteine; Free radicals; Glutathione; Protein disulfides; Protein phosphatases; Thiol oxidation