Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies.
Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies.
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DOI：
10.1021/jacs.6b03983
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发表时间：
2016-07-13
影响因子：
15
通讯作者：
Tycko R
中科院分区：
文献类型：
--
作者：
Bayro MJ;Tycko R
HIV-1衣壳蛋白（CA）形成衣壳壳，将RNA包裹在成熟的HIV-1病毒粒子内。以前的电子显微镜研究表明，衣壳壳主要是CA六聚体的三角形晶格，具有可变的曲率，破坏了平面晶格的理想对称性。成熟的CA晶格依赖于CA二聚化，其通过CA的C-末端结构域（CTD）的螺旋9片段之间的相互作用发生。基于X射线晶体学和多维溶液核磁共振（NMR），已经报道了CTD-CTD二聚化界面的几种高分辨率结构，其中氨基酸侧链构象和螺旋9-螺旋9取向具有显著差异。在基于低温电子显微镜（cryoEM）的管状CA组件的结构模型中[Zhao等人（2013）Nature 497：643 - 646]，二聚化界面基本上是无序的。因此，在非结晶CA组件中的二聚界面结构以及该界面在弯曲晶格内结构有序的程度尚不清楚。在这里，我们描述了固态核磁共振测量的二聚接口在管状CA组件，其中包含一个成熟的病毒粒子的弯曲的三角形晶格，包括定量测量分子间和分子内的距离，使用偶极再耦合技术，固态核磁共振化学位移，和长距离侧链侧链接触。当结合限制从cryoEM研究的螺旋9段之间的距离和取向，固态NMR数据导致一个独特的高分辨率结构的二聚界面中的非结晶晶格的CA管。这些结果表明，CA晶格曲率是不依赖于二聚界面的无序或可变性。这项工作也证明了局部结构测定的可行性内的大型非结晶组装形成的高分子量蛋白质，使用现代固态NMR方法。
The HIV-1 capsid protein (CA) forms the capsid shell that encloses RNA within a mature HIV-1 virion. Previous studies by electron microscopy have shown that the capsid shell is primarily a triangular lattice of CA hexamers, with variable curvature that destroys the ideal symmetry of a planar lattice. The mature CA lattice depends on CA dimerization, which occurs through interactions between helix 9 segments of the C-terminal domain (CTD) of CA. Several high-resolution structures of the CTD-CTD dimerization interface have been reported, based on x-ray crystallography and multidimensional solution nuclear magnetic resonance (NMR), with significant differences in amino acid sidechain conformations and helix 9-helix 9 orientations. In a structural model for tubular CA assemblies based on cryogenic electron microscopy (cryoEM) [Zhao et al. (2013) Nature 497:643–646], the dimerization interface is substantially disordered. The dimerization interface structure in noncrystalline CA assemblies and the extent to which this interface is structurally ordered within a curved lattice have therefore been unclear. Here we describe solid state NMR measurements on the dimerization interface in tubular CA assemblies, which contain the curved triangular lattice of a mature virion, including quantitative measurements of intermolecular and intramolecular distances using dipolar recoupling techniques, solid state NMR chemical shifts, and long-range sidechain-sidechain contacts. When combined with restraints on the distance and orientation between helix 9 segments from the cryoEM study, the solid state NMR data lead to a unique high-resolution structure for the dimerization interface in the noncrystalline lattice of CA tubes. These results demonstrate that CA lattice curvature is not dependent on disorder or variability in the dimerization interface. This work also demonstrates the feasibility of local structure determination within large noncrystalline assemblies formed by high-molecular-weight proteins, using modern solid state NMR methods.
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影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
5.5
作者：
DABBAGH, G;WELIKY, DP;TYCKO, R
通讯作者：
TYCKO, R
DOI：
10.1038/nsb0996-763
发表时间：
1996-09-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Momany, C;Kovari, LC;Rossmann, MG
通讯作者：
Rossmann, MG
影响因子：
2.2
作者：
Bak, M;Rasmussen, JT;Nielsen, NC
通讯作者：
Nielsen, NC
影响因子：
5.4
作者：
Ganser-Pornillos, BK;von Schwedler, UK;Sundquist, WI
通讯作者：
Sundquist, WI