Effect of cellular phospholipid modification on phorbol diester binding.
The influence of cellular lipid composition on the specific binding of [20-3H]phorbol-12,13-dibutyrate to intact human promyelocytic leukemia cells was investigated. Cellular phospholipid composition could be manipulated by culturing cells in serum-free, chemically defined media containing base analogues of phospholipid polar head groups. Human promyelocytic leukemia cells grown in the presence of dimethylethanolamine, monomethylethanolamine, 3-aminopropanol, or isopropylethanolamine assimilated these natural and unnatural base moieties into endogenous phospholipids to the extent that 22 to 52% of the cell glycerophospholipids contained the base analogue. The formation of the phospholipid analogues was accompanied by a pronounced reduction in the levels of intracellular choline and ethanolamine glycerophospholipids. Analogue-supplemented cultures exhibited a reduced growth rate compared to control cells maintained in choline-containing medium. Specific [20-3H ]phorbol-12,13-dibutyrate binding was examined in lipid-altered cells and shown to be markedly higher (approximately 200% of control) in cells grown with dimethyl- or monomethylethanolamine. In contrast, exposure of cells to 3-aminopropanol or isopropylethanolamine resulted in a major reduction in [20-3H]phorbol-12,13-dibutyrate binding. Only minimal changes in nonspecific binding occurred between control and experimental cells. Because phorbol esters are highly membrane targeted, it is possible that phospholipid modification or the resulting changes in membrane organization influence receptor dynamics.
D029742:Caenorhabditis elegans Proteins; D002273:Carcinogens / Q000378:metabolism*; D002352:Carrier Proteins; D002460:Cell Line; D006801:Humans; D007700:Kinetics; D015470:Leukemia, Myeloid, Acute; D015240:Phorbol 12,13-Dibutyrate; D010703:Phorbol Esters / Q000378:metabolism*; D010704:Phorbols / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011493:Protein Kinase C; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011955:Receptors, Drug; D014316:Tritium
D029742:Caenorhabditis elegans Proteins; D002273:Carcinogens; D002352:Carrier Proteins; D010703:Phorbol Esters; D010704:Phorbols; D010743:Phospholipids; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; C083141:phorbol ester binding protein; C027776:phorbol ester receptor; D014316:Tritium; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C