Grunewald R W, Kinne R K
Max-Planck-Institut für Systemphysiologie, Dortmund, Federal Republic of Germany.
Pflugers Arch. 1988 Nov;413(1):32-7. doi: 10.1007/BF00581225.
D-Glucose is an important substrate of energy metabolism and osmolyte synthesis in the renal papillary collecting duct. In order to characterize the cellular entry of D-glucose in this tubular segment, collecting duct cells were isolated from rat kidney papilla and the rate of D-glucose uptake was measured indirectly by monitoring the D-glucose-dependent O2 uptake in the presence of the uncoupler CCCP. D-Glucose uptake was found to be sodium-independent and not sensitive to phlorizin even at a concentration of 10(-3) M. Uptake was, however, completely inhibited by 10(-5) M cytochalasin B and 10(-4) M phloretin. The apparent Ki for cytochalasin B was 1.5 x 10(-6) M and for phloretin 2.0 x 10(-5) M. Studies on the substrate specificity revealed that at 1 mM D-mannose is taken up and metabolized to the same extent as D-glucose. A 50-fold higher concentration of 2-deoxy-D-glucose and 2-amino-2-deoxy-D-glucose inhibited D-glucose uptake completely whereas alpha-methyl-D-glucoside, D-allose, and D-galactose were without effect. Under conditions where D-glucose utilization was maximally stimulated an apparent Km of 1.2 mM and a Vmax of 1 mmol D-glucose/g protein.hour was found for D-glucose uptake. These results indicate that the D-glucose uptake into papillary collecting duct cells is probably mediated by a transport system similar to the one found in basal-lateral membranes of polarized renal, intestinal, and liver cells as well as in nonpolarized fat cells and erythrocytes.
D-葡萄糖是肾乳头集合管能量代谢和渗透溶质合成的重要底物。为了表征该肾小管节段中D-葡萄糖的细胞摄取情况,从大鼠肾乳头分离出集合管细胞,并通过监测解偶联剂CCCP存在下D-葡萄糖依赖性氧气摄取间接测量D-葡萄糖摄取速率。发现D-葡萄糖摄取不依赖于钠,即使在10⁻³ M的浓度下对根皮苷也不敏感。然而,摄取被10⁻⁵ M细胞松弛素B和10⁻⁴ M根皮素完全抑制。细胞松弛素B的表观Ki为1.5×10⁻⁶ M,根皮素为2.0×10⁻⁵ M。底物特异性研究表明,在1 mM时,D-甘露糖的摄取和代谢程度与D-葡萄糖相同。2-脱氧-D-葡萄糖和2-氨基-2-脱氧-D-葡萄糖浓度高50倍时完全抑制D-葡萄糖摄取,而α-甲基-D-葡萄糖苷、D-阿洛糖和D-半乳糖则无作用。在D-葡萄糖利用受到最大刺激的条件下,发现D-葡萄糖摄取的表观Km为1.2 mM,Vmax为1 mmol D-葡萄糖/克蛋白质·小时。这些结果表明,D-葡萄糖摄取到乳头集合管细胞中可能是由一种类似于在极化的肾、肠和肝细胞的基底外侧膜以及非极化的脂肪细胞和红细胞中发现的转运系统介导的。