title: Accumulation of lysophosphatidylinositol in RAW 264.7 macrophage tumor cells stimulated by lipid A precursors.
abstract: N2,O3-Diacylglucosamine 1-phosphate (lipid X), a monosaccharide precursor of Escherichia coli lipid A, was used to stimulate RAW 264.7 macrophage tumor cells, and the effects on macrophage phospholipid metabolism were examined. The addition of E. coli lipid X to the medium of cells that had been uniformly labeled with 32Pi resulted in a 4-8-fold increase in the level of lysophosphatidylinositol. This effect was maximal at 5 microM lipid X. Lysophosphatidylinositol levels reached a maximum 45 min after stimulation, followed by a gradual decline to near normal levels within 2 h. The formation of lysophosphatidylinositol was dependent upon extracellular calcium and was almost completely inhibited when cycloheximide was added at the time of stimulation. The addition of the disaccharide lipid A precursor IVA, commercial lipopolysaccharide (1 microgram/ml), phorbol 12-myristate 13-acetate (10(-7) M), or calcium ionophore A23187 (10(-6) M) to these cells resulted in a similar increase in lysophosphatidylinositol levels, but phosphatidic acid was inactive. The stimulation by IVA and phorbol myristate acetate was blocked by cycloheximide, but the stimulation by lipopolysaccharide was only partially blocked. The stimulation by A23187 was unaffected by cycloheximide. The increase in lysophosphatidylinositol levels might be related to the stimulation of arachidonate release and prostaglandin synthesis that is also observed in cells treated with lipid A precursors. The disaccharide precursor, IVA, was at least 100 times more effective than lipid X at stimulating lysophosphatidylinositol formation and prostaglandin release. The relative ability of lipid X and IVA to stimulate these cells correlated well with their effects on other lipopolysaccharide-responsive systems. Macrophage tumor cells also had the ability to inactivate lipid X by dephosphorylating it.
authors: Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Madison 53706.
authors: R A
authors: RA
authors: Zoeller
authors: P D
authors: PD
authors: Wightman
authors: M S
authors: MS
authors: Anderson
authors: C R
authors: CR
authors: Raetz
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D006017:Glycolipids / Q000494:pharmacology; D008050:Lipid A / Q000494:pharmacology*; D008246:Lysophospholipids / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011453:Prostaglandins / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism*
chemical_list: D006017:Glycolipids; D008050:Lipid A; D008246:Lysophospholipids; D008563:Membrane Lipids; D010743:Phospholipids; D011453:Prostaglandins; C025449:lysophosphatidylinositol; C044283:lipid X; D002118:Calcium