Krupka R M, Devés R
Biochim Biophys Acta. 1980 Jul 16;600(1):228-32. doi: 10.1016/0005-2736(80)90427-7.
A method is described, based on the kinetics of transport, for determining the equilibrium distribution of the carrier site on the inner and outer surfaces of the cell membrane, and this method is applied to the choline carrier of human erythrocytes. This method depends on measurement of flux ratios for both entry and exit, i.e., the transport rates of a low concentration of labeled substrate into a solution which contains either no substrate or a saturating concentration of unlabeled substrate. The concentrations of inward-facing and outward-facing carrier are found to be nearly equal, and therefore the 5-fold difference in choline affinity on the inner and outer surfaces of the membrane cannot be explained by an unequal carrier distribution. It is also shown that both reorientation and dissociation of the carrier-substrate complex are far more rapid than reorientation of the free carrier.
本文描述了一种基于转运动力学的方法,用于确定细胞膜内外表面载体位点的平衡分布,并将该方法应用于人红细胞的胆碱载体。该方法依赖于测量进出通量比,即低浓度标记底物进入不含底物或含有饱和浓度未标记底物的溶液中的转运速率。发现向内和向外的载体浓度几乎相等,因此膜内外表面胆碱亲和力5倍的差异不能用载体分布不均来解释。还表明,载体-底物复合物的重新定向和解离比游离载体的重新定向快得多。