Lees, J. A., Buchkovich, K. J., Marshak, D. R., Anderson, C. W., Harlow, E. (December 1991) The retinoblastoma protein is phosphorylated on multiple sites by human cdc2. EMBO J, 10 (13). pp. 4279-90. ISSN 0261-4189 (Print)0261-4189 (Linking)
Abstract
The retinoblastoma gene product (pRB) is a nuclear phosphoprotein that is thought to play a key role in the negative regulation of cellular proliferation. pRB is phosphorylated in a cell cycle dependent manner, and studies in both actively dividing and differentiated cells suggest that this modification may be essential for cells to progress through the cell cycle. Using tryptic phosphopeptide mapping we have shown that pRB is phosphorylated on multiple serine and threonine residues in vivo and that many of these phosphorylation events can be mimicked in vitro using purified p34cdc2. Using synthetic peptides corresponding to potential cdc2 phosphorylation sites, we have developed a strategy which has allowed the identification of five sites. S249, T252, T373, S807 and S811 are phosphorylated in vivo, and in each case these sites correspond closely to the consensus sequence for phosphorylation by p34cdc2. This and the observation that pRB forms a specific complex with p34cdc2 in vivo suggests that p34cdc2 or a p34cdc2-related protein is a major pRB kinase.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence CDC2 Protein Kinase/*metabolism Cell Cycle Cells, Cultured Chromatography, High Pressure Liquid Electrophoresis, Gel, Two-Dimensional Humans Mass Spectrometry Molecular Sequence Data Peptide Mapping Phosphopeptides/analysis Phosphorylation Precipitin Tests Retinoblastoma Protein/genetics/*metabolism Substrate Specificity Trypsin |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > cdc2 bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | December 1991 |
| Date Deposited: | 16 Dec 2015 20:19 |
| Last Modified: | 16 Dec 2015 20:19 |
| PMCID: | PMC453181 |
| URI: | https://repository.cshl.edu/id/eprint/32133 |
Actions (login required)
 |
Administrator's edit/view item |