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      • SCOPUSKCI등재

        토끼 신장의 근위곱슬세관과 근위곧은세관에 존재하는 Na-α-ketoglutarate Cotransport System의 생화학적 특성

        안도환 ( Do Whan Ahn ),김경룡 ( Kyoung Ryong Kim ),박희석 ( Hee Seok Park ),박양생 ( Yang Saeng Park ) 대한신장학회 2007 Kidney Research and Clinical Practice Vol.26 No.1

        Purpose: alpha-Ketoglutarate (αKG), a Krebs cycle intermediate, is extensively used in the kidney as a fuel substrate and as a counter anion for organic acid secretion. It is known to be taken up by the proximal tubule cells via the brush-border as well as basolateral membranes. We explored biochemical characteristics of the brush-border and basolateral αKG transport systems in pars convoluta and pars recta of the proximal tubule, respectively. Methods: Brush-border and basolateral membrane vesicles (BBMV and BLMV) were isolated from rabbit renal outer cortex and outer medulla by Percoll gradient centrifugation. Vesicular uptake of αKG was determined by rapid Millipore filtration method using α-14[C]KG as a substrate. Results: Both BBMV and BLMV showed a Na-gradient dependent uphill transport of αKG. The systems in both membranes were similarly inhibited by Li and activated by Na (Hill coefficient of 1.4). Kinetic analyses indicated that the Na-αKG cotransporters in the BBMV had a lower substrate affinity as compared with those in the BLMV. The transport systems in BLMVs showed a similar Km but different Vmax between the outer cortex (Km: 34 uM, Vmax: 3.3 nmol/mg protein/10s) and outer medulla (Km: 37, Vmax: 1.8). On the other hand, the systems in BBMVs were different in both Km and Vmax between the outer cortex (Km: 194, Vmax: 3.3) and outer medulla (Km: 89, Vmax: 1.7). Conclusion: The findings suggest that both axial and apical to basolateral heterogeneity of the Na-αKG cotransport system in proximal tubules may be due to a physiological adaptation to efficiently utilize αKG in the kidney.

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