Accession Number : ADA298706

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

Descriptive Note : Annual rept. 1 Jun 94-31 May 95,

Corporate Author : COLD SPRING HARBOR LAB OF QUANTITATIVE BIOLOGY NY

Personal Author(s) : Futcher, A. B.

PDF Url : ADA298706

Report Date : 27 JUN 1995

Pagination or Media Count : 8

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.

Descriptors :   *PROTEINS, *IN VITRO ANALYSIS, *ANTIBODIES, *IN VIVO ANALYSIS, METHODOLOGY, COMMERCE, TWO DIMENSIONAL, BIOCHEMISTRY, ENZYMES, SUBSTRATES, STRAINS(BIOLOGY), YEASTS, GELS, GENETICS, ELECTROPHORESIS, PHOSPHORYLATION.

Subject Categories : Biochemistry
      Medicine and Medical Research

Distribution Statement : APPROVED FOR PUBLIC RELEASE