Swaisgood M, Schindler M
Department of Biochemistry, Michigan State University, East Lansing 48824.
Exp Cell Res. 1989 Feb;180(2):529-36. doi: 10.1016/0014-4827(89)90079-7.
Fluorescence redistribution after photobleaching (FRAP) was utilized to select a "fast" lateral mobility clone from Kirsten murine sarcoma virus-transformed 3T3 (KMSV-3T3) fibroblasts. The clone, E7G1, demonstrated a lateral mobility for membrane wheat germ agglutinin (WGA) and succinylated concanavalin A (sCon A) receptors of (2.1 +/- 1.6) X 10(-9) cm2/s and (2.7 +/- 2.3) X 10(-9) cm2/s, respectively. These mobilities were approximately equivalent to phospholipid mobility (2.8 +/- 1.9 X 10(-9) cm2/s). The fast mobility phenotype is observed when the cells are unattached and spherical. Upon attachment, the mobility decreases to (0.19 +/- 0.19) X 10(-9) cm2/s. In addition, the ability of Con A to initiate global modulation was completely lost in spread as well as spherical cells in the E7G1 fast mobility clone. A comparison of F-actin patterns between untransformed Balb/c fibroblasts and the E7G1-transformed line suggests a correlation between well-developed stress fiber assemblies and the ability to induce global modulation. The fast mobility clone was stable for at least 23 passages.
利用光漂白后的荧光重新分布（FRAP）从 Kirsten 小鼠肉瘤病毒转化的 3T3（KMSV - 3T3）成纤维细胞中选择一个“快速”侧向迁移克隆。该克隆 E7G1 显示膜小麦胚凝集素（WGA）和琥珀酰化伴刀豆球蛋白 A（sCon A）受体的侧向迁移率分别为(2.1±1.6)×10⁻⁹ cm²/s 和(2.7±2.3)×10⁻⁹ cm²/s。这些迁移率大约等同于磷脂的迁移率(2.8±1.9×10⁻⁹ cm²/s)。当细胞未附着且呈球形时观察到快速迁移表型。细胞附着后，迁移率降至(0.19±0.19)×10⁻⁹ cm²/s。此外，在 E7G1 快速迁移克隆的铺展细胞以及球形细胞中，伴刀豆球蛋白 A 引发整体调节的能力完全丧失。未转化的 Balb/c 成纤维细胞与 E7G1 转化细胞系之间 F - 肌动蛋白模式的比较表明，发达的应力纤维组装与诱导整体调节的能力之间存在相关性。快速迁移克隆至少传代 23 次仍保持稳定。
