Abstract
Properties of succinate transport were examined in basolaterat membrane vesicles (BLMV) isolated from rabbit renal cortex. An inwardly directed $Na^+$ gradient stimulated succinate uptake and led to a transient overshoot. $K^+,{\;}Li^+,{\;}Rb^+$ and choline could not substitute for $Na^+$ in the uptake process. The dependence of the initial uptake rate of succinate on $Na^+$ concentration exhibited sigmoidal kinetics, indicating interaction of more than one $Na^+$ with transporter Hill coefficient for $Na^+$ was calculated to be 2.0. The $Na^+-dependent$ succinate uptake was electrogenic, resulting in the transfer of positive charge across the membrane. The succinate uptake into BLMV showed a pH optimum at external pH $7.5{\sim}8.0$, whereas succinate uptake into brush border membrane vesicles (BBMV) did not depend on external pH. Kinetic analysis showed that a Na-dependent succinate uptake in BLMV occurred via a single transport system, with an apparent Km of $15.5{\pm}0.94{\;}{\mu}M$ and Vmax of $16.22{\pm}0.25{\;}nmole/mg{\;}protein/min$. Succinate uptake was strongly inhibited by $4{\sim}5$ carbon dicarboxylates, whereas monocarboxylates and other organic anions showed a little or no effect. The succinate transport system preferred dicarboxylates in trans-configuration (furmarate) over cis-dicarboxylates (maleate). Succinate uptake was inhibited by the anion transport inhibitors DIDS, SITS and furosemide, and $Na^+-coupled$ transport inhibitor harmaline. These results indicate the existence of a $Na^+-dependent$ succinate transport system in BLMV that may be shared by the other Krebs cycle intemediates. This transport system seems to be very similar to the luminal transport system for dicarboxylates.
가토 신장 피질에서 Percoll density gradient방법으로 분리한 basolateral membrane vesicle (BLMV)에서 rapid filtration technique을 이용하여 succinate의 이동 특성을 관찰하였다. $Na^+$은 succinate의 이동을 증가시켜 "overshoot"현상을 보였으며 이러한 효과는 $K^+,{\;}Li^+,{\;}Rb^+,{\;}choline$과 같은 다른 양이온들에 의해 나타나지 않았다. $Na^+$농도변화에 따른 succinate의 이동율은 sigmoid모양을 보였고, $Na^+$에 대한 Hill coefficient는 2.0이었다. soccinate의 이동은 vesicle 내부가 음전압일 때 더욱 증가되었다. BLMV에서 succinate이동은 용액내 pH변화에 따라 영향을 받았으나 brush border membrane vesicle (BBMV)에서는 영향을 받지 않았다. 동력학적 분석결과 succinate의 Km값은 $15.5{\pm}0.94{\;}{\mu}M$이었고 Vmax는 $16.22{\pm}0.25{\;}n{\;}mole/mg{\;}protein/min$이었다. succinate의 이동은 $4{\sim}5$탄소를 가진 dicarboxylate들에 의해 강력하게 억제되었으나 monocarboxylate나 다른 유기음이온들에 의해 영향을 적게 받거나 받지 않았다. succinate의 이동은 DIDS, SITS, furosemide와 같은 음이온 이동 억제제와 harmaline과 같은 $Na^+$ 이동 억제제에 의해 억제되었다. 이들 결과들은 BLMV에서 succinate는 $Na^+$에 의존하여 이동하며 다른 Krebs cycle중간 산물들과 동일한 운반기전을 이용함을 가르킨다. 또한 BLMV에서 succinate의 이동은 그 기질특이성에 있어서 다른 연구자에 의해 보고된 BBMV에서 이동특성과 유사함을 보였다.
(Department of Pharmacology, College of Medicine Pusan National University)