Phosphorylation of endogenous membrane proteins by endogenous protein kinase at the outer surface of Ehrlich cells.
An endogenous protein kinase at the surface of Ehrlich cells has been studied. Using exogenous (gamma32P)ATP as a phosphoryl group donator, a transfer was demonstrated into endogenous acceptor protein(s) as well as to exogenous phosvitin. Seryl- and threonyl-residues isolated from the endogenous and exogenous acceptor protein were found to be labeled. The ratio between the labeled phosphorylserine and phosphorylthreonine was about 3.5:1 for both the endogenous acceptor of the intact cells and the exogenous acceptor. In similar experiments with a membrane preparation from Ehrlich cells, this ratio increased to about 7:1 provided the exogenous acceptor protein was absent. The results were independent of whether 1 X 10(-5) M dibutyryl cyclic AMP was used or not with intact cells and a membrane fraction mainly consisting of vesicles. Whether the regulatory subunit of the membrane-associated protein kinase was in cis- or trans-disposition to the catalytic subunit no binding and dependence of the cyclic nucleotide was observed. Since the purified membrane fraction was considered free from endogenous cyclic AMP, it was concluded that the membrane-associated protein kinase of Ehrlich cells is not dependent on cyclic AMP. The critical role of arginine for the cyclic AMP dependence of the serine-containing residue in the catalytic subunit is discussed.
D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D008297:Male; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism*; D010774:Phosvitin / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism
D009363:Neoplasm Proteins; D013912:Threonine; D000249:Adenosine Monophosphate; D012694:Serine; D003994:Bucladesine; D000255:Adenosine Triphosphate; D010774:Phosvitin; D000242:Cyclic AMP; D011494:Protein Kinases