Construction of covalent membrane protein complexes and high-throughput selection of membrane mimics.
Construction of covalent membrane protein complexes and high-throughput selection of membrane mimics.
复制标题
共价膜蛋白复合物的构建和膜模拟物的高通量选择。
DOI：
10.1021/ja304247f
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发表时间：
2012
影响因子：
15
通讯作者：
Ulmer,TobiasS
中科院分区：
文献类型：
--
作者：
Suk,Jae-Eun;Situ,AlanJ;Ulmer,TobiasS
跨膜螺旋的结合是膜蛋白结构和折叠的基础。TM络合物的结构研究受到络合物稳定性和合适的膜模拟物选择的限制。本文介绍了高效、制备规模构建共价TM复合物和伴随的高通量膜模拟物选择的方法。对于所采用的整合素α ib β3模型体系，该方法确定磷脂单胞体及其特定组成是最佳的膜模拟物。该方法便于通过核磁共振光谱测定结构，如测量以前无法获得的残余偶极耦合和15n弛豫参数。
The association of transmembrane (TM) helices underlies membrane protein structure and folding. Structural studies of TM complexes are limited by complex stability and the often time-consuming selection of suitable membrane mimics. Here, methodology for the efficient, preparative scale construction of covalent TM complexes and the concomitant high-throughput selection of membrane mimics is introduced. For the employed integrin αIIbβ3 model system, the methodology identified phospholipid bicelles, including their specific composition, as the best membrane mimic. The method facilitates structure determination by NMR spectroscopy as exemplified by the measurement of previously inaccessible residual dipolar couplings and15N relaxation parameters.