Protein-bound tyrosine oxidation, nitration and chlorination by-products assessed by ultraperformance liquid chromatography coupled to tandem mass spectrometry

Fecha de publicación: 24/03/2016

Autores de IIS La Fe

María Isabel Torres Cuevas

Autor
Julia Kuligowski

Autor
Consuelo Cháfer Pericás

Autor
Miguel Angel Asensi Miralles

Autor
Elena Cubells Garcia

Autor
Antonio Guillermo Núñez Ramiro

Autor
Máximo Vento Torres

Autor
Justo Javier Escobar Cubiella

Autor

Participantes ajenos a IIS La Fe

Cárcel M
Solberg R
Saugstad OD

Grupos

Perinatología

Leader

Máximo Vento Torres

Abstract

Background: Free radicals cause alterations in cellular protein structure and function. Oxidized, nitrated, and chlorinated modifications of aromatic amino acids including phenylalanine and tyrosine are reliable biomarkers of oxidative stress and inflammation in clinical conditions. Objective: To develop, validate and apply a rapid method for the quantification of known hallmarks of tyrosine oxidation, nitration and chlorination in plasma and tissue proteins providing a snapshot of the oxidative stress and inflammatory status of the organism and of target organs respectively. Material and Methods: The extraction and clean up procedure entailed protein precipitation, followed by protein re-suspension and enzymatic digestion with pronase. An Ultra Performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) method was developed to quantify protein released ortho-tyrosine (o-Tyr), meta-tyrosine (m-Tyr), 3-nitrotyrosine (3NO(2)-Tyr) and 3-chlorotyrosine (3Cl-Tyr) as well as native phenylalanine (Phe) and tyrosine (p-Tyr) in plasma and tissue from a validated hypoxic newborn piglet experimental model. Results: In plasma there was a significant increase in the 3NO(2)-Tyr/p-Tyr ratio. On the other hand m-Tyr/Phe and 3Cl-Tyr/p-Tyr ratios were significantly increased in liver of hypoxic compared with normoxic animals. Although no significant differences were found in brain tissue, a clear tendency to increased ratios was observed under hypoxic conditions. Conclusions: UPLC-MS/MS has proven suitable for the analysis of plasma and tissue samples from newborn piglets. The analysis of biomarkers of protein oxidation, nitration and chlorination will be applied in future studies aiming to provide a deeper insight into the mechanisms of oxidation-derived protein modification caused during neonatal asphyxia and resuscitation. (C) 2016 Elsevier B.V. All rights reserved.

Datos de la publicación

ISSN/ISSNe:: 0003-2670, 1873-4324
Tipo:: Article
Páginas:: 104-110
DOI:: 10.1016/j.aca.2016.01.054
Factor de Impacto:: 1,482 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Métricas

Filiaciones

Torres-Cuevas I:: Instituto de Investigación. Neonatal Research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Kuligowski J:: Instituto de Investigación. Neonatal Research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Cárcel M:: Instituto de Investigación. Neonatal Research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Cháfer-Pericás C:: Instituto de Investigación. Neonatal Research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Asensi M:: Department of Physiology, University of Valencia, Vicent Andrés Estellés s/n, 46100, Burjassot, Spain
Solberg R:: Department of Pediatric Research, Institute for Surgical Research, Oslo University Hospital - Rikshospitalet, Oslo, Norway
Cubells E:: Division of Neonatology, University & Polytechnic Hospital La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Nuñez A:: Division of Neonatology, University & Polytechnic Hospital La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain
Saugstad OD:: Department of Pediatric Research, Institute for Surgical Research, Oslo University Hospital - Rikshospitalet, Oslo, Norway
Vento M:: National Coordinator of the Spanish Maternal and Child Health and Developmental Network, (Retic Red SAMID RD0012/0026), Spain
Escobar J:: Instituto de Investigación. Neonatal Research Group, Health Research Institute La Fe, Avenida Fernando Abril Martorell 101, 46026, Valencia, Spain.