NMR structure of Mistic, a membrane-integrating protein for membrane protein expression
NMR structure of Mistic, a membrane-integrating protein for membrane protein expression
复制标题
DOI：
10.1126/science.1106392
复制
发表时间：
2005-02-25
期刊：
影响因子：
56.9
通讯作者：
Choe, S
中科院分区：
文献类型：
--
作者：
Roosild, TP;Greenwald, J;Choe, S
虽然可溶蛋白质的结构测定已经成为常规，但我们对膜蛋白质的理解一直受到实验瓶颈的限制，无法获得足够的蛋白质产量和有序晶体。Mistic是一种不同寻常的枯草杆菌完整膜蛋白，它可以自动折叠到膜上，绕过细胞转位机制。使用顺磁探针，我们通过核磁共振(核磁共振)光谱确定该蛋白质形成螺旋束，具有令人惊讶的极性面向脂质的表面。更多的实验表明，Mistic可以用于以天然构象高水平生产其他膜蛋白，包括许多以前对细菌难以处理的真核蛋白。表情。
Although structure determination of soluble proteins has become routine, our understanding of membrane proteins has been limited by experimental bottlenecks in obtaining both sufficient yields of protein and ordered crystals. Mistic is an unusual Bacillus subtilis integral membrane protein that folds autonomously into the membrane, bypassing the cellular translocon machinery. Using paramagnetic probes, we determined by nuclear magnetic resonance (NMR) spectroscopy that the protein forms a helical bundle with a surprisingly polar lipid-facing surface. Additional experiments suggest that Mistic can be used for high-level production of other membrane proteins in their native conformations, including many eukaryotic proteins that have previously been intractable to bacterial. expression.