Chapter title |
Segmental Isotopic Labeling of Proteins for NMR Study Using Intein Technology
|
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Chapter number | 9 |
Book title |
Split Inteins
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6451-2_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6449-9, 978-1-4939-6451-2
|
Authors |
Dongsheng Liu, David Cowburn, Liu, Dongsheng, Cowburn, David |
Abstract |
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 72:249-289, 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. |
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