Segmental Isotopic Labeling of Proteins for NMR Study Using Intein Technology
2017
来源专著Methods in Molecular Biology
出版地999 RIVERVIEW DR, STE 208, TOTOWA, NJ 07512-1165 USA
出版者HUMANA PRESS INC
ISSN1064-3745
卷号1495
页码131-145
摘要Segmental isotopic labeling of samples for NMR studies is attractive for large complex biomacromolecular systems, especially for studies of function-related protein-ligand interactions and protein dynamics (Goto and Kay, Curr Opin Struct Biol 10: 585-592, 2000; Rosa et al., Molecules (Basel, Switzerland) 18: 440, 2013; Hiroaki, Expert Opin Drug Discovery 8: 523-536, 2013). Advantages of segmental isotopic labeling include selective examination of specific segment(s) within a protein by NMR, significantly reducing the spectral complexity for large proteins, and allowing for the application of a variety of solution-based NMR strategies. By utilizing intein techniques (Wood and Camarero, J Biol Chem 289: 14512-14519, 2014; Paulus, Annu Rev Biochem 69: 447-496, 2000), two related approaches can generally be used in the segmental isotopic labeling of proteins: expressed protein ligation (Muir, Annu Rev Biochem 72: 249289, 2003) and protein trans - splicing (Shah et al., J Am Chem Soc 134: 11338-11341, 2012). Here, we describe general implementation and latest improvements of expressed protein ligation method for the production of segmental isotopic labeled NMR samples.
关键词Expressed protein ligation Isotopic labeling Intein Structural biology NMR
DOI10.1007/978-1-4939-6451-2_9
收录类别BSCI
语种英语
WOS记录号WOS:000400929800010
通讯作者Liu, Dongsheng
WOS类目Biochemical Research Methods ; Biochemistry & Molecular Biology
WOS研究方向Biochemistry & Molecular Biology
原始文献类型Article ; Book Chapter
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文献类型专著章节
条目标识符https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1590
专题iHuman研究所_特聘教授组_Kurt Wuthrich组
通讯作者Liu, Dongsheng
作者单位
1.ShanghaiTech Univ, iHuman Inst, Shanghai, Peoples R China
2.Albert Einstein Coll Med, Dept Biochem, Bronx, NY 10467 USA
第一作者单位iHuman研究所
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Liu, Dongsheng,Cowburn, David. Segmental Isotopic Labeling of Proteins for NMR Study Using Intein Technology. 999 RIVERVIEW DR, STE 208, TOTOWA, NJ 07512-1165 USA:HUMANA PRESS INC,2017:131-145.
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