A rapid assay for protein kinases phosphorylating small polypeptides and other substrates.
A new and rapid method of protein kinase assay is presented which is suitable for low-molecular-weight substrates, irrespective of their electrophoretic or chromatographic mobility. It depends on the phosphorylation of the substrates with [gamma-32P]ATP, hydrolysis of the pyrophosphate bonds by boiling in 1 N HCl, extraction of 32P with isobutanol-benzene, and measurement of the radioactivity of 32P-labeled phosphoesters in the water phase. The method is shown to be suitable for both tyrosine- and serine-phosphorylating protein kinases.
D000595:Amino Acid Sequence; D000804:Angiotensin II; D000818:Animals; D002364:Caseins; D010455:Peptides / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D014764:Viral Proteins
D002364:Caseins; D010455:Peptides; D010761:Phosphorus Radioisotopes; D014764:Viral Proteins; D000804:Angiotensin II; D011494:Protein Kinases