NMR of Membrane Proteins: Beyond Crystals.
NMR of Membrane Proteins: Beyond Crystals.
复制标题
膜蛋白的核磁共振：超越晶体。
DOI：
10.1007/978-3-319-35072-1_3
复制
发表时间：
2016
影响因子：
--
通讯作者：
Rajesh S
中科院分区：
文献类型：
--
作者：
Rajesh S
膜蛋白对于细胞之间以及细胞的隔室之间的信号、营养物和能量的流动是必不可少的。只有在原子分辨率下定义了所涉及的精确结构、动力学和相互作用后，才能完全理解它们的机制。通过在溶液和固体核磁共振光谱学的进步，这一信息现在是可用的，如稳定的外周和跨膜蛋白的最近的研究所证明的。在这里，我们重点介绍了G蛋白偶联受体、外膜蛋白（例如VDAC）、磷酸肌醇传感器（例如FAPP-1普列克底物蛋白同源结构域）以及包括金属蛋白酶MMP-12在内的酶的最新案例。突出的研究已经导致使用先进的同位素标记策略，增溶系统和设计用于非常高场磁体的NMR实验确定膜相关蛋白的3D结构，动力学性质和相互作用表面。固态核磁共振提供了进一步的见解的结构和多聚体组装的膜蛋白在脂质双层，以及与配体和目标的相互作用。核磁共振在膜结构生物学中的广泛应用所面临的挑战包括需要用于生产具有脆性折叠的同位素标记蛋白质的过表达和纯化系统，以及只有少数昂贵的全氘代去污剂的可用性。可能改变该领域的步骤变化包括聚合物，如苯乙烯马来酸，它完全消除了对去污剂的需求，并允许从细胞或膜直接进行高产率纯化。MODA软件可以促进对NMR的更广泛需求，该软件可以立即预测随后可以通过NMR验证的膜相互作用残基。此外，动态核极化核磁共振仪器的最新发展提供了显着的灵敏度从低摩尔浓度的样品和细胞表面的增强。这些进展说明了目前的能力和未来的潜力，核磁共振膜蛋白结构生物学和配体的发现。
Membrane proteins are essential for the flow of signals, nutrients and energy between cells and between compartments of the cell. Their mechanisms can only be fully understood once the precise structures, dynamics and interactions involved are defined at atomic resolution. Through advances in solution and solid state NMR spectroscopy, this information is now available, as demonstrated by recent studies of stable peripheral and transmembrane proteins. Here we highlight recent cases of G-protein coupled receptors, outer membrane proteins, such as VDAC, phosphoinositide sensors, such as the FAPP-1 pleckstrin homology domain, and enzymes including the metalloproteinase MMP-12. The studies highlighted have resulted in the determination of the 3D structures, dynamical properties and interaction surfaces for membrane-associated proteins using advanced isotope labelling strategies, solubilisation systems and NMR experiments designed for very high field magnets. Solid state NMR offers further insights into the structure and multimeric assembly of membrane proteins in lipid bilayers, as well as into interactions with ligands and targets. Remaining challenges for wider application of NMR to membrane structural biology include the need for overexpression and purification systems for the production of isotope-labelled proteins with fragile folds, and the availability of only a few expensive perdeuterated detergents.Step changes that may transform the field include polymers, such as styrene maleic acid, which obviate the need for detergent altogether, and allow direct high yield purification from cells or membranes. Broader demand for NMR may be facilitated by MODA software, which instantly predicts membrane interactive residues that can subsequently be validated by NMR. In addition, recent developments in dynamic nuclear polarization NMR instrumentation offer a remarkable sensitivity enhancement from low molarity samples and cell surfaces. These advances illustrate the current capabilities and future potential of NMR for membrane protein structural biology and ligand discovery.
登录
查看更多内容
日光细菌光合反应中心两种光化学活性状态的全细胞核磁共振表征。
影响因子：
2.9
作者：
Smitha Surendran Thamarath;A. Alia;Eugenio Daviso;D. Mance;J. Golbeck;J. Matysik
通讯作者：
J. Matysik
影响因子：
16
作者：
Zhou Y;Cierpicki T;Jimenez RH;Lukasik SM;Ellena JF;Cafiso DS;Kadokura H;Beckwith J;Bushweller JH
通讯作者：
Bushweller JH
磷酸肌醇信号传导：比二元更好。
影响因子：
--
作者：
M. Overduin;M. Cheever;T. Kutateladze
通讯作者：
T. Kutateladze
影响因子：
15
作者：
Mao, Jiafei;Nhu-Nguyen Do;Glaubitz, Clemens
通讯作者：
Glaubitz, Clemens
与脂质重构的 Sec 易位子结合的 13C 标记信号肽的动态核极化增强固态 NMR。
影响因子：
15
作者：
Reggie L
通讯作者：
Reggie L