Functional investigations of the human voltage-dependent anion channel 1 by solid-state NMR spectroscopy

膜蛋白可以为各种配体提供通过双层小叶的途径，并且通常以可切换的方式这样做。代谢物在线粒体外膜（MOM）上的运输对于满足活细胞的能量需求至关重要。电压依赖性阴离子通道（VDAC）是MOM中最丰富的蛋白质，是线粒体和细胞质之间代谢物运输的主要途径。283残基hVDAC1已成为决定线粒体介导的细胞凋亡发生的潜在开关。因此，门控机制的知识应该为开发具有促凋亡活性和潜在抗肿瘤药物的小分子提供一个目标。本项目的主要研究目标是hVDAC1在自然环境即脂质双分子层中的结构和功能研究，在脂质双分子层中，hVDAC1是折叠的，在很宽的pH范围内稳定，并且已知具有通道活性。为了实现这些目标，我们计划了大量的固态核磁共振波谱实验，包括二维同核和异核相关光谱以及三维赋值实验。最先进的质子探测将用于进一步改善结构和功能信息。除了VDAC的开放和封闭构象结构外，VDAC与小配体（ATP/NADH）以及生物大分子（如己糖激酶）的相互作用也在计划中。

Membrane proteins can provide pathways through a bilayer leaflet for a variety of ligands and often do so in a switchable fashion. Metabolite transport across the mitochondrial outer membrane (MOM) is essential for fulfilling the energy requirements of living cells. The voltage-dependent anion channel (VDAC) is the most abundant protein of the MOM and is known to be the primary avenue for metabolite traffic between the mitochondrion and cytoplasm. The 283-residue hVDAC1 has emerged as a potential switch determining the onset of mitochondria-relayed apoptosis. Thus, knowledge of the gating mechanism should provide a target for development of small molecules that have pro-apoptotic activity and potential as antitumor agents.The main research objective of this project is the structural and functional study of hVDAC1 in its natural environment, i.e. lipid bilayers, where it is folded, stable over wide pH range, and is known to exhibit channel activity. In order to achieve these goals, a plethora of solid-state NMR spectroscopic experiments is planned, including 2D homo- and heteronuclear correlation spectra as well as 3D assignment experiments. State-of-the-art proton detection will be used to further improve structural and functional information. Besides the structures of VDAC in its open and closed conformation, the interaction of VDAC with small ligands (ATP/NADH) as well as biomacromolecules, such as hexokinase, is planned.