Phosphorylation in membranes of intact human erythrocytes.
Studies of phosphorylation in membranes of intact human erythrocytes were performed by incubating erythrocytes in inorganic [32P]phosphate. Analysis of membrane proteins by polyacrylamide gel electrophoresis showed a pattern of phosphorylation similar to that observed when ghost membranes were incubated with [gamma-32P]ATP. Membrane lipid phosphorylation was also similar in intact cells and ghosts. The most heavily phosphorylated lipid, polyphosphoinositide, was closely associated with glycophorin A, the major erythrocyte membrane sialoglycoprotein obtained when the sialoglycoprotein fraction was isolated by the lithium diiodosalicylate-phenol partition procedure. Only 1 molecule of glycophorin A out of every 100 was found to be phosphorylated, and the phosphate exchange occurred specifically in the COOH-terminal intracellular portion of glycophorin A. These studies show that the human erythrocyte can be used as a model for membrane phosphorylation in an intact cell system.
D000818:Animals; D001386:Azides / Q000494:pharmacology; D001798:Blood Proteins / Q000378:metabolism; D003994:Bucladesine / Q000494:pharmacology; D002417:Cattle; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology; D008565:Membrane Proteins / Q000097:blood*; D009206:Myocardium / Q000201:enzymology; D010446:Peptide Fragments / Q000032:analysis; D010750:Phosphoproteins / Q000097:blood*; D011458:Prostaglandins E / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism; D014357:Trypsin
D001386:Azides; D001798:Blood Proteins; D037102:Lectins; D008565:Membrane Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D011458:Prostaglandins E; D003994:Bucladesine; D011494:Protein Kinases; D014357:Trypsin; D007545:Isoproterenol; D004121:Dimethyl Sulfoxide