An in vitro system to evaluate the scaffold function of the RalA effector protein RalBP1.
The in vitro kinase assay is an invaluable method to identify and characterize kinase substrates and to determine specific site(s) of phosphorylation. However, the use of recombinant proteins does not adequately account for the contribution of accessory proteins, such as scaffolds, that can contribute to the phosphorylation reaction in vivo. We describe here a method to evaluate the contribution of specific scaffold proteins to kinase reactions using a modified in vitro kinase assay.
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, VA, USA.
David F
DF
Kashatus
D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays / Q000379:methods*; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D020725:ral GTP-Binding Proteins / Q000378:metabolism*
D020690:GTPase-Activating Proteins; D011494:Protein Kinases; D020725:ral GTP-Binding Proteins