Unit

Kinase-Catalyzed Biotinylation of Peptides, Proteins, and Lysates

  1. Chamara Senevirathne,
  2. Keith D. Green,
  3. Mary Kay H. Pflum

Published Online: 1 MAR 2012

DOI: 10.1002/9780470559277.ch110228

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

Senevirathne, C., Green, K. D. and Pflum, M. K. H. 2012. Kinase-Catalyzed Biotinylation of Peptides, Proteins, and Lysates. Current Protocols in Chemical Biology. 4:83–100.

Author Information

  1. Department of Chemistry, Wayne State University, Detroit, Michigan

Publication History

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

Abstract

Kinase-catalyzed protein phosphorylation plays an essential role in a variety of biological processes. Methods to detect phosphoproteins and phosphopeptides in cellular mixtures will aid in cell biological and signaling research. Our laboratory recently discovered the utility of γ-modified ATP analogues as tools for studying phosphorylation. Specifically, ATP-biotin can be used for labeling and visualizing phosphoproteins from cell lysates. Because the biotin tag is suitable for protein detection, the biotinylation reaction can be applied to multiple phosphoproteomics applications. Herein, we report a general protocol for labeling phosphopeptides and phosphoproteins in biological samples using kinase-catalyzed biotinylation. Curr. Protoc. Chem. Biol. 4:83-100 © 2012 by John Wiley & Sons, Inc.

Keywords:

  • kinase;
  • ATP-biotin;
  • biotin;
  • phosphoprotein;
  • protein labeling