Abnormal function of protein kinase C in cells having phosphatidylethanolamine-deficient and phosphatidylcholine-excess membranes.
When rat mammary carcinoma 64-24 cells are grown in the absence of ethanolamine, their membrane phospholipid composition changes significantly, becoming phosphatidylethanolamine-deficient and phosphatidylcholine-excess due to a reduced de novo rate of phosphatidylethanolamine synthesis, and growth stops. We have assumed that this membrane phospholipid environment is not suitable for membrane-associated functions. We have previously demonstrated that functions normally stimulated by tumor-promoting phorbol ester, phorbol 12,13-dibutyrate, are not stimulated in ethanolamine-deprived cells, suggesting that function of protein kinase C may be abnormal under the altered membrane environment. In the present study, the behavior of protein kinase C in 64-24 cells grown in the presence and absence of ethanolamine (having normal and phosphatidylethanolamine-deficient/phosphatidylcholine-excess phospholipid) was compared by enzyme assay as well as Western blotting. The results show that the nature of association of protein kinase C to the membrane, which is induced by phorbol ester, is abnormal when cells have the altered membrane phospholipid, and thus argue that membrane phospholipid environment is important in the function of protein kinase C.
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309-0347.
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D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000502:physiology; D006224:Cricetinae; D003600:Cytosol / Q000201:enzymology; D015536:Down-Regulation / Q000502:physiology; D019856:Ethanolamine; D004983:Ethanolamines / Q000494:pharmacology; D008969:Molecular Sequence Data; D015240:Phorbol 12,13-Dibutyrate; D010713:Phosphatidylcholines / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000172:deficiency*; D011493:Protein Kinase C / Q000032:analysis / Q000502:physiology*; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology; D014407:Tumor Cells, Cultured
D004983:Ethanolamines; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D015240:Phorbol 12,13-Dibutyrate; D019856:Ethanolamine; D011493:Protein Kinase C