General Methods for Identifying G1-Phase Substrates of Cdk Protein Kinases.
Abstract
We are using two-dimensional gels in combination with genetic and biochemical techniques to identify G1-phase substrates of cyclin-Cdk complexes. We have optimized protocols for labeling proteins with 32P prior to two-dimensional gel electrophoresis. So far, we have successfully identified one important in vivo substrate, Sic1. We have characterized Sic1, and have shown that phosphorylation of Sic1 by cyclin-Cdk complexes promotes DNA synthesis in yeast. We have made peptide antigens for the purpose of making antibodies against phosphorylated Cdk substrates. Mice have been immunized; their sera are positive; and fusions for monoclonal antibodies are about to be done.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1996
- Accession Number
- ADA323678
Entities
People
- A. B. Futcher
- Daniel R. Marshak