Simulation of NMR data from oriented membrane proteins: practical information for experimental design.
Simulation of NMR data from oriented membrane proteins: practical information for experimental design.
复制标题
定向膜蛋白的 NMR 数据模拟：实验设计的实用信息。
DOI：
10.1016/s0006-3495(93)81215-3
复制
发表时间：
1993
影响因子：
3.4
通讯作者：
Schwonek,JP
中科院分区：
文献类型：
--
作者：
Sanders2nd,CR;Schwonek,JP
对于多孔膜蛋白细菌视紫红质，以及符合几种不同二级结构(α-和3(10)-螺旋以及平行和反平行β-折叠)的理想化跨膜多肽，每个都相对于双层法线有几个倾角，模拟了数百个固体核磁共振偶极偶极耦合和化学位移各向异性。假设使用宏观取向的样本。这些模拟的结果表明：(I)由于偶极耦合对r-3的依赖性，在膜蛋白的定向样品核磁共振研究中，很可能很难成功地执行统一的同位素浓缩策略来产生大量可定量解释的结构测量。(Ii)有许多易于实施的特定同位素标记方案可以产生足以识别相对于双层正常倾斜10度的理想化蛋白质跨膜片段的局部二级结构的数据模式。(Iii)测量13C-、19F-和/或~3H标记的近端残基侧链之间的偶极耦合常数可能被证明是有效的，作为长程三级结构数据约束的途径。
Several hundred solid state NMR dipolar couplings and chemical shift anisotropies were simulated for the polytopic membrane protein, bacteriorhodopsin, and for an idealized transmembrane peptide conforming to several different secondary structures (alpha- and 3(10)-helices and parallel and antiparallel beta-sheets), each at several tilt angles with respect to the bilayer normal. The use of macroscopically oriented samples was assumed. The results of these simulations suggest: (i) Because of the r-3 dependence of dipolar coupling, it is likely to prove difficult to successfully execute uniform isotopic enrichment strategies to generate large numbers of quantitatively interpretable structural measurements in oriented sample NMR studies of membrane proteins. (ii) There are a number of readily implementable specific isotopic labeling schemes which can yield data patterns sufficient to identify local secondary structure for transmembrane segments of idealized proteins which are tilted by < 10 degrees with respect to the bilayer normal. (iii) The measurement of dipolar coupling constants between 13C-, 19F-, and/or 3H-labeled side chains of proximal residues may prove effective as routes to long range tertiary structural data constraints.