Department of Biochemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Adv Exp Med Biol. 2012;992:35-62. doi: 10.1007/978-94-007-4954-2_3.
In this chapter, we summarize the isotopic labeling strategies used to obtain high-quality solution and solid-state NMR spectra of biological samples, with emphasis on integral membrane proteins (IMPs). While solution NMR is used to study IMPs under fast tumbling conditions, such as in the presence of detergent micelles or isotropic bicelles, solid-state NMR is used to study the structure and orientation of IMPs in lipid vesicles and bilayers. In spite of the tremendous progress in biomolecular NMR spectroscopy, the homogeneity and overall quality of the sample is still a substantial obstacle to overcome. Isotopic labeling is a major avenue to simplify overlapped spectra by either diluting the NMR active nuclei or allowing the resonances to be separated in multiple dimensions. In the following we will discuss isotopic labeling approaches that have been successfully used in the study of IMPs by solution and solid-state NMR spectroscopy.
在本章中，我们总结了用于获得生物样品高质量溶液和固态 NMR 谱的同位素标记策略，重点是整合膜蛋白（IMP）。虽然溶液 NMR 用于研究在快速旋转条件下的 IMP，例如在去污剂胶束或各向同性双胶束存在下，但固态 NMR 用于研究脂质囊泡和双层中 IMP 的结构和取向。尽管生物分子 NMR 光谱学取得了巨大进展，但样品的均一性和整体质量仍然是一个需要克服的重大障碍。同位素标记是通过稀释 NMR 活性核或允许共振在多个维度上分离来简化重叠光谱的主要途径。在下面的内容中，我们将讨论已成功用于溶液和固态 NMR 光谱学研究 IMP 的同位素标记方法。
