Screening for Inhibitors of Kinase Autophosphorylation.
Autophosphorylation of kinases influences their conformational state and can also regulate enzymatic activity. Recently, this has become an area of interest for drug discovery. Using Alk2 as an example, we present two protocols - one based on phosphate-binding Alphascreen beads, the other on coupled luminescence measurements of ADP formation - that can be used to screen for inhibitors of autophosphorylation.
Center for Proteomic Chemistry, Novartis Institute for Biomedical Research Basel, Novartis Pharma AG, Postfach, CH-4002, Basel, Switzerland.
Bianca
B
Heedmann
Center for Proteomic Chemistry, Novartis Institute for Biomedical Research Basel, Novartis Pharma AG, Postfach, CH-4002, Basel, Switzerland. martin.klumpp@novartis.com.
Martin
M
Klumpp
D030201:Activin Receptors, Type I / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D010766:Phosphorylation / Q000187:drug effects*; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D000244:Adenosine Diphosphate; C425423:ACVR1 protein, human; D030201:Activin Receptors, Type I
ADP-Glo; Alk2; Alphascreen; Assay development; Autoactivation; Autophosphorylation; High-throughput screening; Kinase; Luminescence