Selective deacylation of 1-acyl-2-arachidonoyl PC and PE in thrombin-stimulated human platelets

Conference · Sat Mar 01 04:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5393852

Purdon, A D; Patelunas, D; Smith, J B

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-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:3
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACYLATION
ALCOHOLS
AMINES
AMMONIUM COMPOUNDS
ARACHIDONIC ACID
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD COAGULATION
BLOOD PLATELETS
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHOLINE
CHROMATOGRAPHY
COAGULANTS
DRUGS
ENZYMES
ESTERS
HEMATOLOGIC AGENTS
HEMOSTATICS
HYDROLASES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPIDS
LIPOTROPIC FACTORS
LIQUID COLUMN CHROMATOGRAPHY
MATERIALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PEPTIDE HYDROLASES
PHOSPHOLIPIDS
QUATERNARY COMPOUNDS
SEPARATION PROCESSES
SERINE PROTEINASES
THIN-LAYER CHROMATOGRAPHY
THROMBIN
TRACER TECHNIQUES
TRITIUM COMPOUNDS