UNIT 14.13 Enrichment and Detection of Tyrosine-Nitrated Proteins
Published Online: 1 AUG 2012
Copyright © 2012 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 AUG 2012
- Published Print: AUG 2012
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.
- mass spectrometry;
- tissue analysis;
- oxidative stress