Work

Covalent capture of kinase-specific phosphopeptides reveals Cdk1-cyclin B substrates

Year: 2008

Type: article

Abstract: We describe a method for rapid identification of protein kinase substrates. Cdk1 was engineered to accept an ATP analog that allows it to uniquely label its substrates with a bio-orthogonal phosphate analog tag. A highly specific, covalent capture-and-release methodology was developed for rapid purification of tagged peptides derived from labeled substrate proteins. Application of this approach to the discovery of Cdk1-cyclin B substrates yielded identification of >70 substrates and phosphorylation sites. Many of these sites are known to be phosphorylated in vivo, but most of the proteins have not been characterized as Cdk1-cyclin B substrates. This approach has the potential to expand our understanding of kinase-substrate connections in signaling networks.

Source: Proceedings of the National Academy of Sciences

DOI: 10.1073/pnas.0708966105

Authors: Justin D. Blethrow, Joseph S. Glavy, David O. Morgan, Kevan M. Shokat

Institutions: Howard Hughes Medical Institute, University of California, San Francisco, Rockefeller University

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green