Open AccessKinase‐Catalyzed Biotinylation of Peptides, Proteins, and Lysates
Author(s) -
Senevirathne Chamara,
Green Keith D.,
Pflum Mary Kay H.
Publication year - 2012
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch110228
Subject(s) - biotinylation , phosphoproteomics , phosphorylation , biotin , biochemistry , kinase , protein kinase a , protein phosphorylation , chemistry , phosphoprotein , proteomics , biology , microbiology and biotechnology , gene
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.