Dalençon F, Rosilio V, Puisieux F, Baszkin A, Wautier J L
Physico-Chimie des Surfaces et Innovation en Pharmacotechnie, URA CNRS 1218. Université Paris-Sud. Châtenay-Malabry, France.
Biochim Biophys Acta. 1996 Aug 16;1302(3):241-8. doi: 10.1016/0005-2760(96)00070-7.
An improved procedure for the direct transfer of membrane proteins from human platelets to liposomes involving the treatment of platelets with linolenic acid was developed. The transfer of platelet proteins to liposomes prepared from the mixture of L-alpha-dimyristoyl-phosphatidylcholine/sphingomyelin in the molar ratio 80/20 appeared to be significantly enhanced compared with liposomes prepared from the same components mixed in other ratios. A wide range of platelet proteins was transferred, the most important being GPIb (170 kDa), GPIIb/IIIa (135 and 110 kDa). GPIV (90 kDa), GPIX (24 kDa) and the serotonin transporter (68 kDa). The recognition interactions between these proteoliposomes and specific protein antibodies clearly indicate that the non-invasive procedure used in this study ensured the reproducible transfer of platelet proteins without essentially altering their original conformation. The obtained results reveal also that the affinity of proteoliposomes to bind paroxetin was virtually the same as that of the native serotonin transporter. These results provide an indication of the possible use of such proteoliposomes as models to study at the molecular level the interaction of these proteins with their ligands.
开发了一种改进的方法，用于将人血小板中的膜蛋白直接转移到脂质体中，该方法涉及用亚麻酸处理血小板。与由相同成分按其他比例混合制备的脂质体相比，由摩尔比为80/20的L-α-二肉豆蔻酰磷脂酰胆碱/鞘磷脂混合物制备的脂质体中，血小板蛋白的转移似乎显著增强。多种血小板蛋白被转移，其中最重要的是糖蛋白Ib（170 kDa）、糖蛋白IIb/IIIa（135和110 kDa）、糖蛋白IV（90 kDa）、糖蛋白IX（24 kDa）和5-羟色胺转运体（68 kDa）。这些蛋白脂质体与特异性蛋白抗体之间的识别相互作用清楚地表明，本研究中使用的非侵入性方法确保了血小板蛋白的可重复转移，而基本上不会改变其原始构象。所获得的结果还表明，蛋白脂质体结合帕罗西汀的亲和力与天然5-羟色胺转运体的亲和力几乎相同。这些结果表明，此类蛋白脂质体有可能作为模型，在分子水平上研究这些蛋白质与其配体的相互作用。
