A tandem chromatographic column method for assaying cAMP-dependent protein kinase and protein kinase C with synthetic peptide substrates.
A method was devised for assaying protein kinases that phosphorylate either Kemptide, such as cAMP-dependent protein kinase, or a glycogen synthase peptide, which is an excellent substrate for protein kinase C. Upon sequential processing of reaction mixtures through tandem columns of cation and anion exchange resins, radioactivity in background samples is nearly nil and the yield of phosphorylated peptides is high. This method reduces labor, radioactivity, enzyme requirements, and costs of assaying protein kinases.
Egan
J J
JJ
Membrane Regulation Section, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
Chang
M K
MK
Londos
C
C
D000273:Adipose Tissue / Q000201:enzymology; D000818:Animals; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods; D003600:Cytosol / Q000201:enzymology; D007202:Indicators and Reagents; D010455:Peptides; D010748:Phosphopeptides / Q000032:analysis*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011493:Protein Kinase C / Q000032:analysis*; D011494:Protein Kinases / Q000032:analysis*; D011865:Radioisotope Dilution Technique; D051381:Rats; D013379:Substrate Specificity
D007202:Indicators and Reagents; D010455:Peptides; D010748:Phosphopeptides; D010761:Phosphorus Radioisotopes; D011494:Protein Kinases; D011493:Protein Kinase C