de Laat S W, van der Saag P T, Elson E L, Schlessinger J
Proc Natl Acad Sci U S A. 1980 Mar;77(3):1526-8. doi: 10.1073/pnas.77.3.1526.
The fluorescence photobleaching recovery method has been used to determine the lateral mobilities of membrane lipids and proteins during the cell cycle of synchronized C1300 mouse neuroblastoma cells (clone Neuro-2A). As probes for lipid mobility, 3,3'-dioctadecylindocarbocyanine iodide and a fluorescein-labeled analog of ganglioside GM1 were used. Membrane proteins were labeled with rhodamine-labeled rabbit antibodies against mouse E14 cells. For both lipid probes the diffusion coefficients reach a minimum in mitosis, increase 2- to 3-fold during G1, remain constant at maximal values during S, and decrease again shortly before mitosis. Membrane proteins also exhibit minimum diffusion coefficients in mitosis, followed by a similar rise in G1. However, as cells proceed through S and G2, the lateral mobility of the membrane proteins gradually decreases. It is argued that lipid mobility is controlled by the fluidity of the membrane lipid matrix whereas protein mobility is governed also by other constraints.
荧光光漂白恢复法已被用于测定同步化的C1300小鼠神经母细胞瘤细胞(克隆Neuro-2A)在细胞周期中膜脂和膜蛋白的侧向迁移率。作为脂类迁移率的探针,使用了3,3'-二辛基吲哚羰花青碘化物和神经节苷脂GM1的荧光素标记类似物。膜蛋白用罗丹明标记的抗小鼠E14细胞兔抗体进行标记。对于两种脂类探针,扩散系数在有丝分裂时达到最小值,在G1期增加2至3倍,在S期保持最大值不变,并在有丝分裂前不久再次下降。膜蛋白在有丝分裂时也表现出最小的扩散系数,随后在G1期有类似的升高。然而,随着细胞进入S期和G2期,膜蛋白的侧向迁移率逐渐降低。有人认为,脂类迁移率受膜脂基质流动性的控制,而蛋白迁移率还受其他限制因素的支配。