A liquid chromatography/mass spectrometry-based method for the selection of ATP competitive kinase inhibitors.
The currently approved kinase inhibitors for therapeutic uses and a number of kinase inhibitors that are undergoing clinical trials are directed toward the adenosine triphosphate (ATP) binding site of protein kinases. The 5'-fluorosulfonylbenzoyl 5'-adenosine (FSBA) is an ATP-affinity reagent that covalently modifies a conserved lysine present in the nucleotide-binding site of most kinases. The authors have developed a liquid chromatography/mass spectrometry-based method to monitor binding of ATP competitive protein kinase inhibitors using FSBA as a nonselective activity-based probe for protein kinases. Their method provides a general, rapid, and reproducible means to screen and validate selective ATP competitive inhibitors of protein kinases.
Khandekar
Sanjay S
SS
Gene Expression and Protein Biochemistry, GlaxoSmithKline, King of Prussia, PA 19406, USA. sanjay.khandekar-1@gsk.com
Feng
Bingbing
B
Yi
Tracey
T
Chen
Susan
S
Laping
Nicholas
N
Bramson
Neal
N
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000737:chemistry; D000345:Affinity Labels / Q000494:pharmacology; D017209:Apoptosis; D001345:Autoradiography; D001665:Binding Sites; D001667:Binding, Competitive; D042846:CDC2-CDC28 Kinases / Q000378:metabolism; D002454:Cell Differentiation; D002853:Chromatography, Liquid / Q000379:methods*; D051357:Cyclin-Dependent Kinase 2; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D010770:Phosphotransferases / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors
D000345:Affinity Labels; D004791:Enzyme Inhibitors; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D042846:CDC2-CDC28 Kinases; D051357:Cyclin-Dependent Kinase 2; D000241:Adenosine