Increasing the accuracy of solution NMR structures of membrane proteins by application of residual dipolar couplings. High-resolution structure of outer membrane protein A
Increasing the accuracy of solution NMR structures of membrane proteins by application of residual dipolar couplings. High-resolution structure of outer membrane protein A
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DOI：
10.1021/ja0608343
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发表时间：
2006-05-31
影响因子：
15
通讯作者：
Bushweller, John H.
中科院分区：
文献类型：
--
作者：
Cierpicki, Tomasz;Liang, Binyong;Bushweller, John H.
膜蛋白的结构测定是溶液核磁共振波谱最具挑战性的应用之一。由于所用的高度氘化蛋白质缺乏距离信息，因此需要新的结构测定方法。在这里，我们证明通过残余偶极耦合（RDC）的精修可以显着提高膜蛋白 OmpA 的结构准确性。带电聚丙烯酰胺凝胶的应用使我们能够获得两种排列并准确测量大量异核偶极耦合。此外，我们已经证明，在细化中使用大量 RDC 可以产生与高分辨率晶体结构主干偏差为 1 埃 rms 的结构。我们对各种数据集的模拟表明，偶极耦合对于获得膜蛋白的准确结构至关重要。
The structure determination of membrane proteins is one of the most challenging applications of solution NMR spectroscopy. The paucity of distance information available from the highly deuterated proteins employed requires new approaches in structure determination. Here we demonstrate that significant improvement in the structure accuracy of the membrane protein OmpA can be achieved by refinement with residual dipolar couplings (RDCs). The application of charged polyacrylamide gels allowed us to obtain two alignments and accurately measure numerous heteronuclear dipolar couplings. Furthermore, we have demonstrated that using a large set of RDCs in the refinement can yield a structure with 1 angstrom rms deviation to the backbone of the high-resolution crystal structure. Our simulations with various data sets indicate that dipolar couplings will be critical for obtaining accurate structures of membrane proteins.