Kaur Anand Ravinderjit, Kanwar Usha, Nath Sanyal Sankar
Department of Biophysics, Panjab University, Chandigarh, India.
Nutr Hosp. 2006 Jan-Feb;21(1):38-46.
Transport characteristics of D-glucose were studied in the microvillous vesicles isolated from the human term placenta. Transport occurred by selective and rapid facilitated diffusion system which was inhibitable by phloretin and HgCl2. The transport was dependent on a transmembrane. Na+-gradient indicating a "secondary active transport" system operating. The transport influx was saturable and the kinetic analysis based on Hanes-Woolf plot produced a kt and Jmax value of 1.2 mM and 34 nmoles.mgprotein(-1).min(-1), respectively. The efflux of D-glucose from the membrane vesicles in a pre-equilibrated assay conditions showed a distinct biphasic pattern differing significantly in the half time efflux. The t1/2 of the fast and slow components was found to be 15 sec and 660 sec, respectively. The transport showed distinct sensitivity to temperature and the Ea values both below and above the transition temperature of 37 degrees C, as calculated from the Arrhenius plot were found to be 7600 and 5472 kCal.mol(-1), respectively. Inhibition studies with a number of sugars for hexose transport pathway showed that the glucose epimers, phosphorylated sugars, and even the disaccharides and the pentose sugars competed effectively with D-glucose. The influx was also inhibited by a number of steroids such as progesterone, 17alpha-hydroxyprogesterone, testosterone and estrogen. Insulin was found to increase glucose transport in a dose- dependent fashion at a concentration of 0.2-1 unit.ml(-1). Ouabain, dinitrophenol and nicotine strongly inhibited D-glucose uptake in the membrane vesicles.
对从足月人胎盘分离出的微绒毛小泡中的D-葡萄糖转运特性进行了研究。转运通过选择性快速易化扩散系统进行,该系统可被根皮素和HgCl2抑制。转运依赖于跨膜Na+梯度,表明存在“继发性主动转运”系统。转运内流是可饱和的,基于Hanes-Woolf图的动力学分析得出的kt和Jmax值分别为1.2 mM和34 nmol·mg蛋白-1·min-1。在预平衡测定条件下,D-葡萄糖从膜小泡中的外流呈现出明显的双相模式,半衰期外流有显著差异。快速和慢速成分的t1/2分别为15秒和660秒。转运对温度表现出明显的敏感性,根据Arrhenius图计算,在37℃转变温度以下和以上的Ea值分别为7600和5472 kCal·mol-1。对多种糖进行的己糖转运途径抑制研究表明,葡萄糖差向异构体、磷酸化糖,甚至二糖和戊糖都能与D-葡萄糖有效竞争。内流也受到多种类固醇的抑制,如孕酮、17α-羟孕酮、睾酮和雌激素。发现胰岛素在0.2-1单位·ml-1的浓度下以剂量依赖方式增加葡萄糖转运。哇巴因、二硝基苯酚和尼古丁强烈抑制膜小泡中D-葡萄糖的摄取。