n-terminally glutamate-substituted analogue of gramicidin a as protonophore and selective mitochondrial uncoupler短杆菌肽的氨基端glutamate-substituted模拟protonophore和选择性线粒体解偶联剂.pdfVIP

N-Terminally Glutamate-Substituted Analogue of Gramicidin A as Protonophore and Selective Mitochondrial Uncoupler 1 1 1 2 Alexandra I. Sorochkina , Egor Y. Plotnikov , Tatyana I. Rokitskaya , Sergei I. Kovalchuk , 1 2 1 1 Elena A. Kotova , Sergei V. Sychev , Dmitry B. Zorov , Yuri N. Antonenko * 1 Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia, 2 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia Abstract Limited uncoupling of oxidative phosphorylation could be beneficial for cells by preventing excessive generation of reactive oxygen species. Typical uncouplers are weak organic acids capable of permeating across membranes with a narrow gap between efficacy and toxicity. Aimed at designing a nontoxic uncoupler, the protonatable amino acid residue Glu was substituted for Val at the N-terminus of the pentadecapeptide gramicidin A (gA). The modified peptide [Glu1]gA exhibited high uncoupling activity in isolated mitochondria, in particular, abolishing membrane potential at the inner mitochondrial membrane with the same or even larger efficacy as gA. With mitochondria in cell culture, the depolarizing activity of [Glu1]gA was observed at concentrations by an order of magnitude lower than those of gA. On the contrary, [Glu1]gA was much less potent in forming proton channels in planar lipid bilayers than gA. Remarkably, at uncoupling concentrations, [Glu1]gA did not alter cell morphology and was nontoxic in MTT test, in contrast to gA showing high toxicity. The difference in the behavior of [Glu1]gA and gA in na