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肾刷状缘膜对三羧酸循环中间产物的钠依赖性转运。对一种电位敏感菁染料荧光的影响。

Na+-dependent transport of tricarboxylic acid cycle intermediates by renal brush border membranes. Effects on fluorescence of a potential-sensitive cyanine dye.

作者信息

Wright S H, Krasne S, Kippen I, Wright E M

出版信息

Biochim Biophys Acta. 1981 Feb 6;640(3):767-78. doi: 10.1016/0005-2736(81)90107-3.

Abstract

The effect of the transport of tricarboxylic acid cycle intermediates on the membrane potential of renal brush border vesicles was studied using fluorescence of the cyanine dye, 3,3'-dipropylthiadicarbocyanine iodide. The behavior of the dye in the preparation was established with valinomycin-induced K+-diffusion potentials; increases in fluorescence were associated with depolarizing conditions. Addition of 1 mm succinate or citrate to membrane/dye suspensions produced transient increases in fluorescence, indicative of a depolarizing event(s) associated with the transport of these substrates. The transient response in fluorescence was Na+ dependent, of greater magnitude under Na+-gradient as compared to Na+-equilibrium conditions, and was a saturable function of substrate concentration. The specificity of the fluorescence response was identical to that obtained from studies of the competitive inhibition of succinate transport by tricarboxylic acid cycle intermediates and analogs We conclude that the major tricarboxylic acid cycle intermediates are transported via a common Na+-dependent transport system in renal brush border membranes.

摘要

利用花青染料3,3'-二丙基硫代二碳菁碘化物的荧光,研究了三羧酸循环中间体的转运对肾刷状缘小泡膜电位的影响。通过缬氨霉素诱导的K⁺扩散电位确定了制剂中染料的行为;荧光增加与去极化条件相关。向膜/染料悬浮液中添加1 mM琥珀酸盐或柠檬酸盐会导致荧光短暂增加,表明与这些底物转运相关的去极化事件。荧光的短暂反应依赖于Na⁺,与Na⁺平衡条件相比,在Na⁺梯度下幅度更大,并且是底物浓度的饱和函数。荧光反应的特异性与通过三羧酸循环中间体和类似物对琥珀酸盐转运的竞争性抑制研究获得的特异性相同。我们得出结论,主要的三羧酸循环中间体通过肾刷状缘膜中一种常见的Na⁺依赖性转运系统进行转运。

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