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 labelling proteins with 32P prior to two-dimensional gel electrophoresis. So far, we have successfully identified and characterized one important in vivo substrate, Sic 1. Other workers have shown that an analogous substrate, p27, is important in human cells. More recently, we have identified and started to characterize two more substrates, Mcm3 and Cdc54. We have made peptide antigens for the purpose of making antibodies against phosphorylated Cdk substrates. Monoclonal antibodies possibly reactive against phospho-Ser-Pro are now being tested for their exact specificity.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1997
- Accession Number
- ADA332674
Entities
People
- A. B. Futcher
- Daniel R. Marshak