Use of a double-headed peptide substrate to study the specificity of cAMP-dependent protein kinase and posphorylase kinase.
A peptide containing 2 seryl residues, (1)Leu(2)Ser(3)Tyr(4)Arg(5)Aly(6)Tyr(7)Ser(8)Leu, was chemically synthesized and used as a substrate for phosphorylase kinase and cyclic AMP-dependent protein kinase. The sequence, TryArgGlyTyr, makes up a beta turn in the native protein. Phosphorylase kinase was found to phosphorylate specifically seryl residue2 and protein kinase seryl residue7. Km and Vmax values were obtained and compared with natural substrates. The differences in the specificity of the two enzymes might be explained by a different requirement for organized structure. As a working hypothesis, it is suggested the results could be explained if the two enzymes interacted with seryl residues at different sides of a beta turn.
D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000494:pharmacology; D004789:Enzyme Activation; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D009132:Muscles / Q000201:enzymology; D009842:Oligopeptides / Q000138:chemical synthesis*; D010764:Phosphorylase Kinase / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
D009842:Oligopeptides; D000242:Cyclic AMP; D011494:Protein Kinases; D010764:Phosphorylase Kinase