Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Bologna, Italy.
Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Bologna, Italy.
Biochimie. 2014 Feb;97:128-37. doi: 10.1016/j.biochi.2013.10.002. Epub 2013 Oct 11.
The antibiotic oligomycin is known to inhibit mitochondrial F-type ATP synthases. The antibiotic inhibits both ATP synthesis and hydrolysis by blocking the H(+) translocation through FO which is coupled to the catalytic activity of F1. The amphiphilic organotin tri-n-butyltin (TBT), a known mitochondrial poison, can penetrate into biological membranes and covalently bind to electron-donor atoms of biomolecules such as sulfur. This study aims at exploring the mechanism(s) involved in the enzyme desensitization to oligomycin which occurs at concentrations >1 μM TBT. This poorly known effect of TBT, which only appeared at temperatures above the break in the Arrhenius plot of the enzyme activity, was found to be accompanied by the oxidation of isolated thiol groups of the mitochondrial complex. The oligomycin sensitivity was restored by the reducing agents glutathione and dithioerythritol and not influenced by antioxidants. The whole of data is consistent with the hypothesis that thiol oxidation is caused by TBT covalent binding to cysteine residues in a low-affinity site on FO and not by other possible oxidative events. According to this putative model, the onset of tin-sulfur bonds would trigger conformational changes and weaken the oligomycin interaction with FO.
寡霉素是一种已知的抑制线粒体 F 型 ATP 合酶的抗生素。该抗生素通过阻止与 F1 的催化活性偶联的 FO 中的 H+转运来抑制 ATP 的合成和水解。亲脂性有机锡三丁基锡(TBT)是一种已知的线粒体毒物,可穿透生物膜并与生物分子(如硫)的供电子原子共价结合。本研究旨在探索寡霉素酶脱敏的机制,这种现象发生在浓度 >1 μM TBT 时。TBT 的这种鲜为人知的作用仅在酶活性的 Arrhenius 图的断裂温度以上出现,与分离的线粒体复合物的巯基氧化有关。还原剂谷胱甘肽和二硫苏糖醇可恢复寡霉素的敏感性,而抗氧化剂则不受影响。所有数据都与假设一致,即巯基氧化是由 TBT 与 FO 上低亲和力部位的半胱氨酸残基发生共价结合引起的,而不是由其他可能的氧化事件引起的。根据这个假设模型,锡硫键的形成会引发构象变化,从而削弱寡霉素与 FO 的相互作用。