Powers M F, Beavis A D
Department of Pharmacology, Medical College of Ohio, Toledo 43699-0008.
J Biol Chem. 1991 Sep 15;266(26):17250-6.
The inner membrane of liver and heart mitochondria possesses an anion uniport pathway, known as the inner membrane anion channel (IMAC). IMAC is inhibited by matrix Mg2+, matrix H+, N,N'-dicyclohexycarbodiimide, mercurials and amphiphilic amines such as propranolol. Most of these agents react with a number of different mitochondrial proteins and, therefore, more selective inhibitors have been sought. In this paper, we report the discovery of a new class of inhibitors, triorganotin compounds, which block IMAC completely. One of the most potent, tributyltin (TBT) inhibits malonate uniport via IMAC 95% at 0.9 nmol/mg. The only other mitochondrial protein reported to react with triorganotins, the F1F0ATPase, is inhibited by about 0.75 nmol/mg. The potency of inhibition of IMAC increases with hydrophobicity in the sequence trimethyltin much less than triethyltin much less than tripropyltin less than triphenyltin less than tributyltin; which suggests that the binding site is accessible from the lipid bilayer. It has long been established that triorganotins are anionophores able to catalyze Cl-/OH- exchange; however, TBT is able to inhibit Cl- and NO3- transport via IMAC at doses below those required to catalyze rapid rates of Cl-/OH- exchange. Consistent with previous reports, the data indicate that about 0.8 nmol of TBT per mg of mitochondrial protein is tightly bound and not available to mediate Cl-/OH- exchange. We have also shown that the mercurials, p-chloromercuribenzene sulfonate and mersalyl, which only partially inhibit Cl- and NO3- transport can increase the IC50 for TBT 10-fold. This effect appears to result from a reaction at a previously unidentified mercurial reactive site. The inhibitory dose is also increased by raising the pH and inhibition by TBT can be reversed by S2- and dithiols but not by monothiols.
肝脏和心脏线粒体的内膜具有一种阴离子单向转运途径,称为内膜阴离子通道(IMAC)。IMAC受到基质Mg2+、基质H+、N,N'-二环己基碳二亚胺、汞剂以及亲脂性胺类(如普萘洛尔)的抑制。这些试剂大多会与多种不同的线粒体蛋白发生反应,因此一直在寻找更具选择性的抑制剂。在本文中,我们报告了一类新型抑制剂——三有机锡化合物的发现,这类化合物能完全阻断IMAC。其中最有效的一种,三丁基锡(TBT)在0.9 nmol/mg时通过IMAC抑制丙二酸单向转运达95%。据报道,唯一另一种能与三有机锡发生反应的线粒体蛋白,F1F0ATP酶,在约0.75 nmol/mg时受到抑制。IMAC的抑制效力随疏水性增加,顺序为三甲基锡远小于三乙基锡小于三丙基锡小于三苯基锡小于三丁基锡;这表明结合位点可从脂质双层进入。长期以来已经确定,三有机锡是能够催化Cl-/OH-交换的阴离子载体;然而,TBT能够在低于催化快速Cl-/OH-交换所需剂量的情况下抑制通过IMAC的Cl-和NO3-转运。与先前的报道一致,数据表明每毫克线粒体蛋白约0.8 nmol的TBT紧密结合,无法介导Cl-/OH-交换。我们还表明,仅部分抑制Cl-和NO3-转运的汞剂对氯汞苯磺酸盐和汞撒利,可使TBT的IC50增加10倍。这种效应似乎是由在一个先前未确定的汞反应位点发生的反应导致的。通过提高pH值也会增加抑制剂量,TBT的抑制作用可被S2-和二硫醇逆转,但不能被单硫醇逆转。