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首页> 外文期刊>Journal of Molecular Biology >Direct evidence for the cooperative unfolding of cytochrome c in lipid membranes from H-H-2 exchange kinetics
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Direct evidence for the cooperative unfolding of cytochrome c in lipid membranes from H-H-2 exchange kinetics

机译：H-H-2交换动力学中脂质膜中细胞色素c协同展开的直接证据

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The interaction of cytochrome c (cyt c) with anionic lipid membranes is known to disrupt the tightly packed native structure of the protein. This process leads to a lipid-inserted denatured state, which retains a nativelike cc-helical structure but lacks any specific tertiary interactions. The structural and dynamic properties of cyt c bound to vesicles containing an anionic phospholipid (DOPS) were investigated by amide H-H-2 exchange using two-dimensional NMR spectroscopy and electrospray ionisation mass spectrometry. The H-H-2 exchange kinetics of the core amide protons in cyt c, which in the native protein undergo exchange via an uncorrelated EX2 mechanism, exchange in the lipid vesicles via a highly concerted global transition that exposes these protected amide groups to solvent. The lack of pH dependence and the observation of distinct populations of deuterated and protonated species by mass spectrometry confirms that exchange occurs via an EX1 mechanism with a common rate of 1(+/-0.5) h(-1) which reflects the rate of transition from the lipid-inserted state, H', to an unprotected conformation, D', associated with the lipid interface. (C) 2000 Academic Press. [References: 44]
机译：已知细胞色素c（cyt c）与阴离子脂质膜的相互作用会破坏蛋白质的紧密堆积的天然结构。该过程导致脂质插入的变性状态，其保留天然的cc-螺旋结构，但缺乏任何特定的三级相互作用。通过使用二维NMR光谱和电喷雾电离质谱的酰胺H-H-2交换研究了与包含阴离子磷脂（DOPS）的囊泡结合的Cyt c的结构和动力学性质。 cyt c中核心酰胺质子的H-H-2交换动力学，其在天然蛋白中通过不相关的EX2机理进行交换，在脂质囊泡中通过高度协调的全局过渡进行交换，从而使这些受保护的酰胺基暴露于溶剂中。缺乏pH依赖性以及通过质谱观察到的氘化和质子化物种的不同种群证实，交换是通过EX1机制发生的，其共同速率为1（+/- 0.5）h（-1），反映了过渡速率从脂质插入状态H'到与脂质界面相关的未保护构象D'。 （C）2000学术出版社。 [参考：44]

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