Plagemann P G, Wohlhueter R M
Biochim Biophys Acta. 1982 Jun 14;688(2):505-14. doi: 10.1016/0005-2736(82)90362-5.
The kinetics of hypoxanthine transport were measured in hypoxanthine phosphoribosyltransferase-deficient Novikoff cells by rapid kinetic techniques applying both zero-trans and equilibrium exchange protocols. The data indicate operation of a simple carrier with directional symmetry and equal mobility when substrate loaded and empty. Zero-trans influx and efflux were about equivalent and so were zero-trans influx and equilibrium exchange flux. The apparent Michaelis-Menten constant and maximum velocity were about 500 microM and 100 pmol/s per microliter cell H2O, respectively. The time courses of accumulation of radioactively labeled hypoxanthine at a concentration above the Michaelis-Menten constant differed noticeably in zero-trans and equilibrium exchange mode, but computer simulations showed that the difference is predicted by the symmetrical carrier model and does not reflect trans-stimulation.
采用零转运和平衡交换实验方案的快速动力学技术,测定了次黄嘌呤磷酸核糖基转移酶缺陷型诺维科夫细胞中次黄嘌呤转运的动力学。数据表明,存在一种简单载体,其具有方向对称性,且底物装载和空载时的迁移率相等。零转运流入和流出大致相等,零转运流入和平衡交换通量也大致相等。表观米氏常数和最大速度分别约为500微摩尔和每微升细胞水100皮摩尔/秒。在高于米氏常数的浓度下,放射性标记次黄嘌呤积累的时间进程在零转运和平衡交换模式下有明显差异,但计算机模拟表明,这种差异可由对称载体模型预测,并不反映反式刺激。