General Methods for Identifying G1 Substrates of the Cdk Protein Kinases.

Abstract

We are using two-dimensional gels in combination with genetic and biochemical techniques to identify Gi-phase substrates of cyclin-Cdk complexes. We have optimized protocols for labelling proteins with 32P prior to two-dimensional gel electrophoresis. Yeast strains have been constructed bearing six different cyclins expressed from a GAL promoter, to allow inducible phosphorylation of the desired substrates. A commercial antibody has been identified and obtained which may specifically immmunoprecipitate phosphorylated forms of the desired substrates. Putative substrates have been examined in vitro. Together, these accomplishments provide a good platform from which to identify real, in vivo substrates in the near future.

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Document Details

Document Type
Technical Report
Publication Date
Jun 27, 1995
Accession Number
ADA298706

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  • A. B. Futcher

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  • Cold Spring Harbor Laboratory

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  • Biology

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