Enrichment and detection of tyrosine-nitrated proteins

Frank Dekker, Nicolas Abello, Rosalina Wisastra, Rainer Bischoff, Frank Dekker

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

Nitrotyrosine is a post-translationally modified amino acid with distinctly different properties than tyrosine or any other of the genetically encoded amino acids. Detecting proteins containing nitrotyrosine is the first step towards a better understanding of the role of nitrotyrosine in health and disease. Moreover, quantifying the extent of nitrotyrosine and determining its location in a protein forms the basis for a better understanding of the effect of tyrosine nitration on biological function. Described in this unit is a method to detect tyrosine-nitrated proteins in tissue sections and on western blots after creating a fluorescent complex between aminotyrosine, salicylaldehyde, and Al(3+). In addition, an approach is detailed for labeling aminotyrosine with biotin to enrich peptides from complex samples. Both methods require reduction of nitrotyrosine to aminotyrosine, which can be achieved with sodium dithionite or hemin plus dithiothreitol.

Original languageEnglish
Pages (from-to)14.13.1 - 14-13-19
Number of pages19
JournalCurrent Protocols in Protein Science
Volume69
Issue number1
DOIs
Publication statusPublished - Aug-2012

Keywords

  • Animals
  • Biotin
  • Blotting, Western
  • Fluorescent Dyes
  • Humans
  • Immunochemistry
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Models, Molecular
  • Oxidation-Reduction
  • Proteins
  • Staining and Labeling
  • Tyrosine

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