title: Phosphorylation of chloroform soluble compounds in plasma membranes of human epidermoid carcinoma A431 cells.
abstract: This study investigated a possible role for the epidermal growth factor (EGF) receptor protein tyrosine kinase in phosphoinositide metabolism with plasma membrane vesicles from human epidermoid carcinoma (A431) cells. We found a novel chloroform-soluble product radiolabeled with [gamma-32P]ATP that did not migrate from the origin in the thin layer system designed to separate the phosphoinositides, appeared as a single band of Mr = 3500 on polyacrylamide gels in the presence of dodecyl sulfate, had an ultraviolet absorbance spectrum with a maximum at 275 nm and stained with Coomassie dye. Based on these properties this phosphorylation product is referred to as a proteolipid. The 32P label was not detected in phosphotyrosine [Tyr(P)], phosphoserine [Ser(P)] or phosphothreonine [Thr(P)] and was lost during acid or base hydrolysis. Phosphorylation of proteolipid was increased significantly by EGF, whereas phosphorylation of phosphatidic acid was decreased and labeling of phosphoinositides was unaffected. Thus, it appears that in A431 membranes the EGF receptor/kinase does not utilize phosphatidylinositol as a substrate, but does phosphorylate a membrane proteolipid.
authors.forename: D L
authors.initials: DL
authors.lastname: Brautigan
authors.forename: P
authors.initials: P
authors.lastname: Randazzo
authors.forename: C
authors.initials: C
authors.lastname: Shriner
authors.forename: J N
authors.initials: JN
authors.lastname: Fain
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002725:Chloroform; D002855:Chromatography, Thin Layer; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D012995:Solubility
chemical_list: D008563:Membrane Lipids; D010716:Phosphatidylinositols; D010761:Phosphorus Radioisotopes; D004815:Epidermal Growth Factor; D002725:Chloroform; D000255:Adenosine Triphosphate