Fujii T, Tamura A, Yamane T
J Biochem. 1985 Nov;98(5):1221-7. doi: 10.1093/oxfordjournals.jbchem.a135388.
14C-Labeled phosphatidylcholine (PC) and lysophosphatidylcholine (lysoPC) species with two homologous saturated acyl chains and of a saturated acyl chain of various lengths, respectively, were each incorporated into the outer leaflet of the membrane lipid bilayer of intact human erythrocytes, and the transbilayer movement into the inner leaflet during incubation at 37 degrees C of the lipid-loaded erythrocytes was followed. The labeled PC and lysoPC molecules present in the outer leaflet were extracted with egg-yolk PC liposome suspension and BSA solution, respectively, and the amount which moved into the inner leaflet during the incubation was measured by determining the residual amount of the labeled lipid in the membrane. Translocation of lysoPC molecules was also measured by assaying the decrease in the amount of the added labeled lysoPC in the membrane during the incubation on the basis of the previously reported fact that lysoPC molecules are all converted metabolically to PC or glycerylphosphorylcholine plus fatty acid as soon as they are translocated from the outer to the inner leaflet. Every lipid tested showed significant transbilayer movement during the course of the incubation for up to 10 h. With the C8, C10, and C12 species of PC the rate of the transbilayer movement increases with decreasing acyl chain length. The same is true with the C14, C16, and C18-lysoPC species.
分别将具有两条同源饱和酰基链和不同长度饱和酰基链的14C标记的磷脂酰胆碱(PC)和溶血磷脂酰胆碱(lysoPC)种类掺入完整人红细胞膜脂双层的外小叶中,并跟踪脂质负载的红细胞在37℃孵育期间向内膜小叶的跨膜运动。外小叶中存在的标记PC和lysoPC分子分别用蛋黄PC脂质体悬浮液和BSA溶液提取,并通过测定膜中标记脂质的残留量来测量孵育期间进入内小叶的量。基于先前报道的事实,即lysoPC分子一旦从外小叶转运到内小叶就会全部代谢转化为PC或甘油磷酰胆碱加脂肪酸,通过测定孵育期间膜中添加的标记lysoPC量的减少来测量lysoPC分子的转运。所测试的每种脂质在长达10小时的孵育过程中均显示出明显的跨膜运动。对于PC的C8、C10和C12种类,跨膜运动速率随酰基链长度的减小而增加。C14、C16和C18-lysoPC种类也是如此。