McKean M L, Smith J B, Silver M J
J Biol Chem. 1982 Oct 10;257(19):11278-83.
Arachidonic acid and other fatty acids are taken up by human platelets from plasma and incorporated into membrane phospholipids. However, little is known about the mechanism and specificity of the various steps of fatty acid insertion into phospholipid. Previous findings from this laboratory have shown that the incorporation of radioactive C20-unsaturated fatty acids (arachidonic, 8,11,14-eicosatrienoic, and 5,8,11,14,17-eicosapentaenoic) into the phospholipids of "resting"p platelets is more rapid than that of the radioactive C16- and C18-saturated and unsaturated fatty acids. We now provide evidence that human platelet microsomes contain acyl-CoA:1-acyl-sn-glycero-3-phosphocholine acyltransferase. The enzyme preparation has a pH optimum of 7.0. Activity is insensitive to 1 mM EDTA and is inhibited 37% by 1 mM Ca2+ and 20% by 1 mM Mg2+. Maximal activity is observed at 100 microM 1-acyl lysophosphatidylcholine at several concentrations of fatty acyl-CoA esters. Apparent Km values from 1.05 to 5.70 microM were obtained for saturated and unsaturated fatty acyl group donors in the presence of 100 microM 1-acyl lysophosphatidylcholine as fatty acyl group acceptor. Comparison of the apparent Vmax values showed that unsaturated CoA esters were transferred more rapidly to 1-acyl lysophosphatidylcholine than saturated CoA esters. Oleate, linoleate, and arachidonate, the major unsaturated fatty acids in platelet phosphatidylcholine, were transferred at similar rates. 8,11,14-eicosatrienoate was transferred about two times faster than these three fatty acyl groups. The data indicate that the incorporation of unsaturated fatty acids into phosphatidylcholine by human platelets occurs via reacylation of 1-acyl lysophosphatidylcholine.
花生四烯酸和其他脂肪酸可被人体血小板从血浆中摄取并掺入膜磷脂中。然而,关于脂肪酸插入磷脂各个步骤的机制和特异性,人们了解甚少。本实验室先前的研究结果表明,放射性C20不饱和脂肪酸(花生四烯酸、8,11,14-二十碳三烯酸和5,8,11,14,17-二十碳五烯酸)掺入“静止”血小板的磷脂中的速度比放射性C16和C18饱和及不饱和脂肪酸更快。我们现在提供证据表明,人体血小板微粒体含有酰基辅酶A:1-酰基-sn-甘油-3-磷酸胆碱酰基转移酶。该酶制剂的最适pH值为7.0。其活性对1 mM EDTA不敏感,1 mM Ca2+可抑制37%,1 mM Mg2+可抑制20%。在几种脂肪酸辅酶A酯浓度下,当1-酰基溶血磷脂酰胆碱浓度为100 microM时观察到最大活性。在以100 microM 1-酰基溶血磷脂酰胆碱作为脂肪酰基受体的情况下,饱和和不饱和脂肪酰基供体的表观Km值在1.05至5.70 microM之间。表观Vmax值的比较表明,不饱和辅酶A酯比饱和辅酶A酯更快地转移到1-酰基溶血磷脂酰胆碱上。油酸、亚油酸和花生四烯酸是血小板磷脂酰胆碱中的主要不饱和脂肪酸,它们的转移速率相似。8,11,14-二十碳三烯酸的转移速度比这三个脂肪酰基快约两倍。数据表明,人体血小板将不饱和脂肪酸掺入磷脂酰胆碱是通过1-酰基溶血磷脂酰胆碱的再酰化作用实现的。