Equilibrium exchanges in the range of 2-40 mM-3-O-methyl glucose at 16 degrees C suggested that the half-saturation concentration for exchange was 22 mM and that the maximum velocity (Vmax) was ca. 149 mmol l-1 min-1. 2. Initial rates of exchange influx from 1, 2, 4 and 8 mM into 76 mM solution gave a half-saturation value of 3.6 mM and a Vmax of 122 mmol-1 min-1. 3. The non-transportable inhibitor 4,6-O-ethylidene-alpha-D-glucopyranose (ethylidene glucose) acting on the outside of the cells inhibited 3-O-methyl glucose exchanges at 16 degrees C with an inhibition constant (KI) of ca. 11 mM. 4. Sen-Widdas exit experiments gave the half-saturation for 3-O-methyl glucose at 16 degrees C as only ca. 2 mM and the KI for ethylidene glucose as ca. 4 mM. 5. Efflux inhibitions by ethylidene glucose are satisfactorily predicted by the asymmetric carrier kinetics of Regen & Tarpley (1974) when using the parameters derived from the exchange experiments but not with parameters from Sen-Widdas exits. 6. Uphill transfer by counterflow experiments and Sen-Widdas exits cannot be fitted by the Regen and Tarpley kinetics (using the same parameters) unless the kinetics are modified to provide for an extra exchange element which replaces some of the net exit component in the equations. 7. At present the modification to the kinetics is only possible in computer simulations and data handling, but with it the fit to experimental results is good. The nature of the modification is described and in the light of it a revised interpretation of the significance of the Km derived from Sen-Widdas exits is discussed.
摘要
16℃下,2 - 40 mM的3 - O - 甲基葡萄糖的平衡交换表明,交换的半饱和浓度为22 mM,最大速度(Vmax)约为149 mmol·l⁻¹·min⁻¹。2. 从1、2、4和8 mM流入76 mM溶液的交换初始速率给出的半饱和值为3.6 mM,Vmax为122 mmol⁻¹·min⁻¹。3. 作用于细胞外的非转运抑制剂4,6 - O - 亚乙基 - α - D - 吡喃葡萄糖(亚乙基葡萄糖)在16℃下抑制3 - O - 甲基葡萄糖的交换,抑制常数(KI)约为11 mM。4. 森 - 威达斯流出实验表明,16℃下3 - O - 甲基葡萄糖的半饱和值仅约为2 mM,亚乙基葡萄糖的KI约为4 mM。5. 当使用从交换实验得出的参数时,亚乙基葡萄糖的流出抑制可通过Regen和Tarpley(1974)的不对称载体动力学得到满意预测,但使用森 - 威达斯流出实验的参数则不然。6. 逆流实验和森 - 威达斯流出实验的上坡转运无法用Regen和Tarpley动力学(使用相同参数)拟合,除非对动力学进行修改,以提供一个额外的交换元素,该元素在方程中取代一些净流出成分。7. 目前,动力学的修改仅在计算机模拟和数据处理中可行,但有了它,对实验结果的拟合很好。描述了修改的性质,并据此讨论了对从森 - 威达斯流出实验得出的Km意义的修订解释。