Charged gels as orienting media for measurement of residual dipolar couplings in soluble and integral membrane proteins
Charged gels as orienting media for measurement of residual dipolar couplings in soluble and integral membrane proteins
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DOI：
10.1021/ja046054g
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发表时间：
2004-12-15
影响因子：
15
通讯作者：
Bushweller, JH
中科院分区：
文献类型：
--
作者：
Cierpicki, T;Bushweller, JH
膜蛋白残留偶极偶联的测量将极大地提高通过溶液核磁共振波谱获得的结构的质量。虽然在实现膜结合多肽的比对方面已经取得了一些成功，但在实现功能膜蛋白的比对方面取得的成功非常有限。在这里，我们证明了带电的基于聚丙烯酰胺的共聚物适合于获得在洗涤剂胶束中重组的膜蛋白的弱排列。通过改变共聚物组成，我们制备了正离子、两性离子和负电荷凝胶，这些凝胶在低浓度下非常稳定，可以用于在核磁共振管中通过压缩获得弱排列。将该方法应用于DPC胶束中完整膜蛋白OmpA的测定。
Measurement of residual dipolar couplings for membrane proteins will dramatically improve the quality of the structures obtainable by solution NMR spectroscopy. While there has been some success in achieving alignment of membrane-bound peptides, there has been very limited success in achieving alignment for functional membrane proteins. Herein, we demonstrate that charged polyacrylamide-based copolymers are suitable for obtaining weak alignment of membrane proteins reconstituted in detergent micelles. Varying the copolymer compositions, we prepared positively, zwitterionic, and negatively charged gels that are very stable at low concentration and can be used for obtaining weak alignment by compression in an NMR tube. Application of this method is demonstrated for the integral membrane protein OmpA in DPC micelles.