Selective deacylation of 1-acyl-2-arachidonoyl PC and PE in thrombin-stimulated human platelets
Previously the authors have shown that uptake and stimulated release of /sup 3/H-arachidonate (AA) in human platelets involves mainly 1-acyl-2-arachidonoyl phospholipids. To determine deacylation of molecular species of 1-acyl-2-arachidonoyl phosphatidylcholine (PC) and phosphatidylethanolamine (PE), phospholipid was extracted by the method of Bligh and Dyer from cells (10/sup 9//ml) stimulated or not by thrombin (THR, 5 U/ml .37/sup 0/C, 5 min). Total PC and PE were isolated by thin-layer chromatography (TLC) and converted to 1-radyl-2-acyl glycerobenzoates. The 1,2 diacyl glycerobenzoates were separated from other subclasses by TLC. Individual molecular species of 1,2 diacyl glycerobenzoates were resolved by reverse phase HPLC. Mass (O.D./sub 230/) and /sup 3/H-AA radioactivity (i.e. specific activity) were determined on-line, and changes in individual molecular species could be deduced. Significant deacylation of all 1-acyl-2-arachidonoyl PC and PE molecular species occurred with no apparent deacylation in any non-AA-containing molecular species of 1,2 diacyl PC/PE. At 5 minutes the net deacylation of 1-acyl-2-arachidonoyl PC and PE was approximately 15 and 5 nanomoles, respectively/10/sup 9/ cells. These results indicate that selective deacylation of arachidonoyl-containing molecular species compared to non-arachidonoyl-containing molecular species of PC/PE occurs in THR-stimulated cells. This suggests certain AA-containing phospholipids are compartmentalized with and susceptible to, the action of phospholipase A/sub 2/.
- Research Organization:
- Temple Univ., Philadelphia, PA
- OSTI ID:
- 5393852
- Report Number(s):
- CONF-8604222-; TRN: 86-028304
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
Similar Records
Transfer of arachidonate from phosphatidylcholine to phosphatidylethanolamine and triacylglycerol in guinea pig alveolar macrophages
Relative turnover of (/sup 3/H)arachidonic acid and (/sup 14/C)eicosapentaenoic acid in stimulated human platelets
Related Subjects
AMINES
ACYLATION
BLOOD PLATELETS
BLOOD COAGULATION
CHOLINE
ARACHIDONIC ACID
LIQUID COLUMN CHROMATOGRAPHY
PHOSPHOLIPIDS
THIN-LAYER CHROMATOGRAPHY
THROMBIN
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ALCOHOLS
AMMONIUM COMPOUNDS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
COAGULANTS
DRUGS
ENZYMES
ESTERS
HEMATOLOGIC AGENTS
HEMOSTATICS
HYDROLASES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPIDS
LIPOTROPIC FACTORS
MATERIALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PEPTIDE HYDROLASES
QUATERNARY COMPOUNDS
SEPARATION PROCESSES
SERINE PROTEINASES
550201* - Biochemistry- Tracer Techniques