Solid-state NMR study of proteorhodopsin in the lipid environment: Secondary structure and dynamics
Solid-state NMR study of proteorhodopsin in the lipid environment: Secondary structure and dynamics
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DOI：
10.1016/j.bbamem.2009.09.011
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发表时间：
2009-12-01
影响因子：
3.4
通讯作者：
Ladizhansky, Vladimir
中科院分区：
文献类型：
--
作者：
Shi, Lichi;Lake, Evelyn M. R.;Ladizhansky, Vladimir
视紫红质是一种典型的视网膜结合光驱动质子泵，具有七螺旋结构，广泛分布于海洋和淡水细菌中。最近我们已经表明，可以在脂质结合状态下制备绿色蛋白视紫红质（GPR），其在具有不同反向标记模式的样品中给出良好分辨的魔角自旋（MAS）NMR光谱。在这里，我们提出了3D和4D连续化学位移分配确定通过实验进行的均匀C-13，N-15标记的样品。这些实验提供了153个残基的分配，在跨膜区具有特别高的密度（类似于74%的残基）。任务的范围允许详细检查的二级结构和动态GPR。特别是，我们目前的实验证据的流动性的蛋白质的末端和A-B，C-D，和F-G环，后者可能耦合到GPR离子转运功能。(C)2009爱思唯尔有限公司版权所有。
Proteorhodopsins are typical retinal-binding light-driven proton pumps of heptahelical architecture widely distributed in marine and freshwater bacteria. Recently. we have shown that green proteorhodopsin (GPR) can be prepared in a lipid-bound state that gives well-resolved magic angle spinning (MAS) NMR spectra in samples with different patterns of reverse labelling. Here, we present 3D and 4D sequential chemical shift assignments identified through experiments conducted on a uniformly C-13,N-15-labelled sample. These experiments provided the assignments for 153 residues, with a particularly high density in the transmembrane regions (similar to 74% of residues). The extent of assignments permitted a detailed examination of the secondary structure and dynamics in GPR. In particular, we present experimental evidence of mobility of the protein's termini and of the A-B, C-D, and F-G loops, the latter being possibly coupled to the GPR ion-transporting function. (C) 2009 Elsevier B.V. All rights reserved.