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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Acknowledgments
DL acknowledges ShanghaiTech University and Shanghai Municipal Government for financial support; Supported by NIH grants GM47042 (DC).
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Liu, D., Cowburn, D. (2017). Segmental Isotopic Labeling of Proteins for NMR Study Using Intein Technology. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_9
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