Schlessinger J, Koppel D E, Axelrod D, Jacobson K, Webb W W, Elson E L
Proc Natl Acad Sci U S A. 1976 Jul;73(7):2409-13. doi: 10.1073/pnas.73.7.2409.
We report measurements of the lateral mobility of fluorescent labeled concanavalin A receptor complexes on the plasma membrane of cultured myoblasts of rat. Transport rates were measured by observing the recovery of fluorescence in a small region of the cell surface initially photobleached irreversibly by an intense, focused laser light pulse. Under different conditions we measured effective diffusion coefficients of the receptor complexes in the range 8 x 10(-12) less than D less than 3 x 10(-11) cm2/sec which is two orders of magnitude lower than we found for a fluorescent lipid probe, D approximately (8 +/- 3) x 10(-9) cm2/sec. This large difference and the presence of apparently immobile concanavalin A receptors suggests that factors beyond the fluoidity of the phospholipid bilayer membrane matrix control the rate of lateral transport of the complexes. Effective mobilities of the complexes decrease with increases in the valence, dose, and occupation time of the lectin on the membrane. These properties imply an aggregation of the lectin-receptor complexes. Mobilities are not influenced by azide, colchicine or preincubation at low temperature. Cytochalasin B and low temperatures, during the time of measurement, decrease the lateral transport rate.
我们报告了对大鼠培养成肌细胞质膜上荧光标记的伴刀豆球蛋白A受体复合物侧向迁移率的测量结果。通过观察细胞表面一个小区域内荧光的恢复来测量转运速率,该区域最初被强聚焦激光脉冲不可逆地光漂白。在不同条件下,我们测量了受体复合物的有效扩散系数,其范围为8×10⁻¹²<D<3×10⁻¹¹cm²/秒,这比我们用荧光脂质探针测得的结果低两个数量级,荧光脂质探针的D约为(8±3)×10⁻⁹cm²/秒。这种巨大差异以及明显不可移动的伴刀豆球蛋白A受体的存在表明,除了磷脂双分子层膜基质的流动性之外,还有其他因素控制着复合物的侧向转运速率。复合物的有效迁移率随着凝集素在膜上的价态、剂量和占据时间的增加而降低。这些特性意味着凝集素 - 受体复合物发生了聚集。迁移率不受叠氮化物、秋水仙碱或低温预孵育的影响。在测量期间,细胞松弛素B和低温会降低侧向转运速率。