Meddings J B, DeSouza D, Goel M, Thiesen S
University of Calgary, Alberta, Canada.
J Clin Invest. 1990 Apr;85(4):1099-107. doi: 10.1172/JCI114541.
Both transport function and microvillus membrane physical properties evolve as the enterocyte matures and migrates up the crypt-villus axis. We isolated enriched fractions of villus tip, mid-villus, and crypt enterocytes from which microvillus membrane vesicles were prepared. Using this material we characterized the alterations that occur in microvillus membrane fluidity as the rabbit enterocyte matures and correlated these with kinetic studies of glucose transport. With increasing maturity the microvillus membrane becomes more rigid due to both an increase in the cholesterol/phospholipid ratio and alterations in individual phospholipid subclasses. Maximal rates of glucose transport were greatest in microvillus membrane vesicles prepared from mature cells. However, the glucose concentration producing half-maximal rates of transport (Km) was significantly lower in crypt microvillus membrane vesicles, suggesting that a distinct glucose transporter existed in crypt enterocytes. This distinction disappeared when differences between membrane lipid environments were removed. By fluidizing villus-tip microvillus membrane vesicles, in vitro, to levels seen in the crypt microvillus membrane, we observed a reduction in the Km of this transport system. These data suggest that the kinetic characteristics of the sodium-dependent glucose transporter are dependent upon its local membrane environment.
随着肠上皮细胞成熟并沿隐窝-绒毛轴向上迁移,其转运功能和微绒毛膜物理特性都会发生变化。我们从绒毛顶端、绒毛中部和隐窝肠上皮细胞中分离出富集部分,用于制备微绒毛膜囊泡。利用这些材料,我们表征了随着兔肠上皮细胞成熟,微绒毛膜流动性发生的变化,并将这些变化与葡萄糖转运的动力学研究相关联。随着成熟度增加,由于胆固醇/磷脂比率增加以及个别磷脂亚类的变化,微绒毛膜变得更加坚硬。在从成熟细胞制备的微绒毛膜囊泡中,葡萄糖转运的最大速率最高。然而,在隐窝微绒毛膜囊泡中,产生半数最大转运速率(Km)的葡萄糖浓度显著更低,这表明在隐窝肠上皮细胞中存在一种独特的葡萄糖转运体。当消除膜脂质环境差异时,这种差异消失。通过在体外将绒毛顶端微绒毛膜囊泡的流动性调节至隐窝微绒毛膜中的水平,我们观察到该转运系统的Km降低。这些数据表明,钠依赖性葡萄糖转运体的动力学特性取决于其局部膜环境。