Edidin M, Wei T
J Cell Biol. 1982 Nov;95(2 Pt 1):458-62. doi: 10.1083/jcb.95.2.458.
We have used fluorescence photobleaching and recovery (FPR) to measure the lateral diffusion of mouse H-2 antigens, labeled with fluorescent Fab fragments, in the membrane of cl 1d fibroblasts. Diffusion coefficients, D, vary more than 20-fold from cell to cell, though they vary no more than twofold when measured at different points on a single cell. The fraction of H-2 antigens mobile, R, also varies from cell to cell, and no lateral diffusion of H-2 antigens can be detected in approximately 20% of the cells examined. Treatment of cells with NaCN + NaF, reducing their levels of ATP reduces the proportion of cells in which no lateral diffusion can be detected. The maximum values of D seen in poisoned cells are less than those in controls. Treatment of cells with the divalent inophore, A23187, greatly increases the proportion of cells in which diffusion of H-2 is rapid, D greater than 2 x 10(-9) cm2 s-1. The data obtained on diffusion by FPR can be replotted in the form of an experiment in which lateral diffusion of H-2 antigens is measured in a population of heterokaryons. There is good agreement between this transformation and actual data on heterokaryons. Thus the two methods appear to measure the same transport process.
我们利用荧光光漂白恢复技术(FPR)来测量用荧光Fab片段标记的小鼠H-2抗原在cl 1d成纤维细胞膜中的侧向扩散。扩散系数D在不同细胞间变化超过20倍,而在单个细胞的不同点测量时,其变化不超过两倍。可移动的H-2抗原的比例R在不同细胞间也有所不同,在所检测的细胞中,约20%的细胞未检测到H-2抗原的侧向扩散。用NaCN + NaF处理细胞,降低其ATP水平,可减少未检测到侧向扩散的细胞比例。中毒细胞中观察到的D的最大值低于对照组。用二价离子载体A23187处理细胞,极大地增加了H-2扩散迅速(D大于2×10⁻⁹ cm² s⁻¹)的细胞比例。通过FPR获得的关于扩散的数据可以重新绘制为在一群异核体中测量H-2抗原侧向扩散的实验形式。这种转换与异核体的实际数据有很好的一致性。因此,这两种方法似乎测量的是相同的转运过程。