The logic and design of analog-sensitive kinases and their small molecule inhibitors.
Analog-sensitive AS Kinase technology allows for rapid, reversible, and highly specific inhibition of individual engineered kinases in cells and in mouse models of human diseases. The technique consists of two parts: a kinase containing a space-creating mutation in the ATP-binding pocket and a bulky ATP-competitive small molecule inhibitor that complements the shape of the mutant ATP pocket. This strategy enables dissection of phospho-signaling pathways, elucidation of the physiological function of individual kinases, and characterization of the pharmacology of clinical-kinase inhibitors. Here, we present an overview of AS technology and describe a stepwise approach for generating AS Kinase mutants and identifying appropriate small molecule inhibitors. We also describe commonly encountered technical obstacles and provide strategies to overcome them.
Howard Hughes Medical Institute and Department of Cellular & Molecular Pharmacology, University of California, San Francisco, California, USA.
Michael S
MS
Lopez
Howard Hughes Medical Institute and Department of Cellular & Molecular Pharmacology, University of California, San Francisco, California, USA.
Joseph I
JI
Kliegman
Howard Hughes Medical Institute and Department of Cellular & Molecular Pharmacology, University of California, San Francisco, California, USA.
Kevan M
KM
Shokat
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D015195:Drug Design; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D015202:Protein Engineering; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013499:Surface Properties
D008024:Ligands; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D011494:Protein Kinases; C478705:purine-analog-sensitive kinase, human
Analog-sensitive; Chemical Genetics; Kinase