Tamura A, Yoshikawa K, Fujii T, Ohki K, Nozawa Y, Sumida Y
Biochim Biophys Acta. 1986 Feb 27;855(2):250-6. doi: 10.1016/0005-2736(86)90171-9.
TEMPO-phosphatidylcholine (PC) spin probes which have homologous saturated acyl chains of 10, 12, 14 and 16 carbon atoms, were synthesized as analogues of PC. Transfer of TEMPO-PCs from liposomal membrane to the ghost membrane of human erythrocyte and transverse diffusion of TEMPO-PCs within the membrane of intact erythrocytes were determined by measurement of spontaneous increase and decrease in signal amplitude of an anisotropic triplet spectrum, due to dilution of the label by natural phospholipid of the membrane and reduction of the label by the cytoplasmic content of the erythrocyte, respectively. TEMPO-PC molecules in TEMPO-PC liposomes, except dipalmitoyl TEMPO-PC, were rapidly incorporated into the ghost membrane by incubation at 37 degrees C; the PC having shorter acyl chains was transferred faster. The cytoplasmic content of the erythrocyte rapidly reduced the nitroxide radical of the spin probe. The central peak height of ESR signal was once increased by incorporation of TEMPO-PC into the erythrocyte membrane and then was spontaneously decreased during further incubation at 37 degrees C. This decrease indicates that PC molecules traverse from the outer to the inner layer of the membrane lipid bilayer. The decrease of signal amplitude was faster with PC of shorter acyl chain. These findings suggest that both transfer between membranes and transverse diffusion in the membrane may be favored to the PC species with shorter acyl chains.
合成了具有10、12、14和16个碳原子的同源饱和酰基链的TEMPO - 磷脂酰胆碱(PC)自旋探针作为PC的类似物。通过测量各向异性三重态光谱信号幅度的自发增加和减少来确定TEMPO - PCs从脂质体膜到人类红细胞鬼膜的转移以及TEMPO - PCs在完整红细胞膜内的横向扩散,这分别是由于膜的天然磷脂对标记物的稀释和红细胞细胞质成分对标记物的还原所致。除二棕榈酰TEMPO - PC外,TEMPO - PC脂质体中的TEMPO - PC分子在37℃孵育时迅速掺入鬼膜;酰基链较短的PC转移得更快。红细胞的细胞质成分迅速还原了自旋探针的氮氧自由基。ESR信号的中心峰高在TEMPO - PC掺入红细胞膜时曾一度增加,然后在37℃进一步孵育期间自发下降。这种下降表明PC分子从膜脂双层的外层穿越到内层。酰基链较短的PC信号幅度下降得更快。这些发现表明,膜间转移和膜内横向扩散可能都有利于酰基链较短的PC种类。