Comparison of diglyceride production from choline-containing phosphoglycerides in human neutrophils stimulated with N-formylmethionyl-leucylphenylalanine, ionophore A23187 or phorbol 12-myristate 13-acetate
- 15 September 1992
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 286 (3), 693-699
- https://doi.org/10.1042/bj2860693
Abstract
The turnover of choline-containing phosphoglycerides (PC) in response to agonist stimulation is well documented in human neutrophils. We have now compared the enzymic pathways of N-formylmethionyl-leucylphenylalanine (fMLP)-, A23187- and phorbol-12-myristate 13-acetate (PMA)-induced diglyceride (DG) and phosphatidic acid (PA) generation in these cells. In order to distinguish between phospholipase C- and D-mediated PC breakdown, human neutrophils were radiolabelled with 1-O-[3H]alkyl-2-acyl-glycero-3-phosphocholine and stimulated in the presence of ethanol or propranolol. The addition of 0.5% ethanol to the incubation mixture resulted in the production of phosphatidylethanol, indicative of phospholipase D activation, in response to all three stimuli. Concomitant with phosphatidylethanol formation was a partial block of PA production. The production of DG was also partially blocked by addition of ethanol. Propranolol (200 microM) was also used to assess the contributions of phospholipases C and D toward DG generation. Inhibition of PA phosphohydrolase by propranolol resulted in the complete abolition of DG generation when neutrophils were stimulated with fMLP. In contrast, propranolol only partially inhibited DG generation in response to A23187 and PMA. These results suggested that DG production in response to fMLP stimulation is mediated via the activation of phospholipase D, whereas A23187- or PMA-induced DG generation may involve more than one pathway. However, examination of the water-soluble choline metabolites produced indicated that phospholipase D was responsible for the production of PA and DG in response to all three stimuli.Keywords
This publication has 36 references indexed in Scilit:
- Bradykinin stimulates phosphodiesteratic cleavage of phosphatidylcholine in cultured endothelial cellsBiochemical and Biophysical Research Communications, 1988
- The phosphatidylcholine pathway of diacylglycerol formation stimulated by phorbol diesters occurs via phospholipase D activationFEBS Letters, 1988
- Activation of phospholipase D by chemotactic peptide in HL-60 granulocytesBiochemical and Biophysical Research Communications, 1988
- Inositol lipids and cell proliferationBiochimica et Biophysica Acta (BBA) - Reviews on Cancer, 1987
- Direct evidence for phorbol ester-stimulated accumulation of diacylglycerol derived from phosphatidyicholineCarcinogenesis: Integrative Cancer Research, 1987
- Phorbol ester induced 1,2-diacylglycerol accumulation and protein phosphorylation in C3H/10T1/2 mouse embryo cellsCarcinogenesis: Integrative Cancer Research, 1985
- Chemical and biochemical aspects of platelet activating factor: A novel class of acetylated ether‐linked choline‐phospholipidsMedicinal Research Reviews, 1985
- Release of arachidonic acid from 1-ALKYL-2-ACYL-sn-glycero-3-phosphocholine, a precursor of platelet-activating factor, in sat alveolar macrophagesBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1984
- 1-O-Alkyl-2-acyl-sn-glycero-3-phosphocholine: A novel source of arachidonic acid in neutrophils stimulated by the calcium ionophore A23187Biochemical and Biophysical Research Communications, 1983
- A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATIONCanadian Journal of Biochemistry and Physiology, 1959