Unit

UNIT 14.13 Enrichment and Detection of Tyrosine-Nitrated Proteins

  1. Frank Dekker1,
  2. Nicolas Abello2,
  3. Rosalina Wisastra1,
  4. Rainer Bischoff2

Published Online: 1 AUG 2012

DOI: 10.1002/0471140864.ps1413s69

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Dekker, F., Abello, N., Wisastra, R. and Bischoff, R. 2012. Enrichment and Detection of Tyrosine-Nitrated Proteins. Current Protocols in Protein Science. 69:14.13:14.13.1–14.13.19.

Author Information

  1. 1

    Pharmaceutical Gene Modulation, Groningen Research Institute of Pharmacy, University of Groningen, The Netherlands

  2. 2

    Analytical Biochemistry, Groningen Research Institute of Pharmacy, University of Groningen, The Netherlands

Publication History

  1. Published Online: 1 AUG 2012
  2. Published Print: AUG 2012

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 Al3+. 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. Curr. Protoc. Protein Sci. 69:14.13.1-14.13.19. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • nitrotyrosine;
  • nitration;
  • fluorescence;
  • mass spectrometry;
  • tissue analysis;
  • oxidative stress