Determination of molecular motion in membranes using periodic pattern photobleaching.
Determination of molecular motion in membranes using periodic pattern photobleaching.
复制标题
使用周期性模式光漂白测定膜中的分子运动。
DOI：
10.1073/pnas.75.6.2759
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发表时间：
1978
影响因子：
11.1
通讯作者：
H. Mcconnell
中科院分区：
文献类型：
--
作者：
B. Smith;H. Mcconnell
荧光磷脂探针在二肉豆蔻酰磷脂酰胆碱定向多层中的横向扩散通过观察膜光漂白后平行条纹的周期性图案中荧光的重新分布来测量。发现荧光脂质的扩散常数 D 在单斜晶相中在 9.6 度时的 1.5 X 10(-11) cm2/sec 和 22.5 度时的 2.0 X 10(-10) cm2/sec 之间变化。对 Lbeta' 和 Pbeta' 相中二棕榈酰磷脂酰胆碱脂质体的初步研究产生了 10(-11) cm2/sec 量级的扩散常数。这些数据与半抗原致敏脂质体膜补体激活率的早期讨论相关[Brûlet, P. 和 McConnell, H. M. (1976) Proc.国家。阿卡德。科学。美国73,2977--2981; Parce, J. W.、Henry, N. 和 McConnell, H.M. (1978) 程序。国家。阿卡德。科学。美国 75, 1515--1518)]。我们还使用这种方法来研究与小鼠 EL-4 肿瘤细胞结合的荧光抗体的运动。图案光漂白技术的优点是细胞或脂质体翻译对测量没有重大不利影响，可以检测和表征某些非扩散运动，并且可以用照相方式记录扩散或其他运动。
The lateral diffusion of a fluorescent phospholipid probe in oriented multibilayers of dimyristoylphosphatidylcholine has been measured by observing the redistribution of fluorescence after photobleaching of the membranes in a periodic pattern of parallel stripes. The diffusion constant D of the fluorescent lipid was found to vary between 1.5 X 10(-11) cm2/sec at 9.6 degrees and 2.0 X 10(-10) at 22.5 degrees in the monoclinic phase. Preliminary studies of dipalmitoylphosphatidylcholine liposomes in the Lbeta' and Pbeta' phases yielded diffusion constants of the order of 10(-11) cm2/sec. These data are relevant to earlier discussions of the rate of complement activation by hapten-sensitized liposomal membranes [Brûlet, P. and McConnell, H. M. (1976) Proc. Natl. Acad. Sci. USA 73,2977--2981; Parce, J. W., Henry, N. and McConnell, H.M. (1978) Proc. Natl. Acad. Sci. USA 75, 1515--1518)]. We have also used this method to study the motion of fluorescent antibodies bound to murine EL-4 tumor cells. Pattern photobleaching techniques have the advantages that cellular or liposomal translation has no major adverse effect on the measurements, that certain nondiffusive motions can be detected and characterized, and that diffusive or other motions can be recorded photographically.