Detection of distinct pools of the adapter protein p130CAS using a panel of monoclonal antibodies.
Dynamic protein interactions are thought to play an important role in regulating a wide variety of signal transduction pathways. Adapter molecules that contribute to the assembly and disassembly of these protein complexes are likely to play a critical role in the regulation of these pathways. The function of one such adapter molecule, p130CAS (CAS), has been implicated in signaling pathways involving cell growth, adhesion, and differentiation. We report here the isolation and characterization of a panel of monoclonal antibodies that specifically recognize CAS. These antibodies are proving to be invaluable molecular reagents for defining the expression, phosphorylation, binding partners, and ultimately the function of CAS with respect to cell signaling. In addition to their utility as conventional reagents for protein isolation, a subset of these antibodies has also proven to be a sensitive tool for distinguishing between different tyrosine-phosphorylated pools of CAS in the cell. Because tyrosine phosphorylation of CAS provides a dynamic means with which to regulate protein-protein interactions, these antibodies may thus serve as molecular reagents that can discern the protein binding potential of CAS. Collectively, the antibodies described in this report provide the means with which to define specific roles for CAS in cell signaling that have been otherwise difficult to establish.
Bouton
A H
AH
Department of Microbiology and Cancer Center, University of Virginia, Charlottesville 22908, USA.
Burnham
M R
MR
D000818:Animals; D000911:Antibodies, Monoclonal; D002645:Chickens; D050719:Crk-Associated Substrate Protein; D000882:Haplorhini; D006801:Humans; D051379:Mice; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000032:analysis*; D010766:Phosphorylation; D011506:Proteins; D051381:Rats; D016160:Retinoblastoma Protein / Q000032:analysis*; D050717:Retinoblastoma-Like Protein p130
D000911:Antibodies, Monoclonal; C493841:BCAR1 protein, human; C493842:Bcar1 protein, mouse; C493843:Bcar1 protein, rat; D050719:Crk-Associated Substrate Protein; D010750:Phosphoproteins; D011506:Proteins; D016160:Retinoblastoma Protein; D050717:Retinoblastoma-Like Protein p130