By Wolfgang J. Baumann
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285, Institut Pasteur, 75015 Paris, France The metabolism of 1-acyl-2-acetyl-sn–glycero-3-phosphocholine (1-acyl-PAF), a naturally occurring analogue of platelet activating factor (PAF), was investigated in rabbit platelets. Our studies showed that 1-acyl-[3H]PAF (1-palmitoyl-2-acetyl-sn-glycero-3-phospho [N-methyl-3H]choline) was converted by platelets into phosphatidyl[3H]choline ([3H]PC) in a time-dependent fashion. The formation of [3H]PC occurred at a rate similar to that observed when lyso-[3H]PC (palmitoyl-sn-glycero-3phospho[N-methyl-3H]choline) was used as substrate.
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Plastic silica gel plates for thin-layer chromatography (TLC) and organic solvents were from Merck, Ltd. (Darmstadt, West Germany). Preparation of radioactive 1-acyl-PAF. [3H]PC (50 μCi) was hydrolyzed by 30-min incubation with 10 μg of bee venom PLA2 in Tyrode’s buffer containing 2 mM Ca2+. The lyso-[3H]PC produced by this hydrolysis was then converted into 1-acyl-[3H]PAF (1-palmitoyl-2-acetyl-sn- 993 1-ACYL-PAF METABOLISM IN RABBIT PLATELETS glycero-3-phospho[N-methyl-3H]choline) by acetylation according to the method of Wientzek et al.