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Split Inteins pp 131–145Cite as

Segmental Isotopic Labeling of Proteins for NMR Study Using Intein Technology

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1495))

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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Authors and Affiliations

  1. iHuman Institute, ShanghaiTech University, 99 Haike Road, Pudong, Shanghai, 201203, China

    Dongsheng Liu

  2. Department of Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA

    David Cowburn

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  1. Dongsheng Liu
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  2. David Cowburn
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Correspondence to David Cowburn .

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Editors and Affiliations

  1. Institute of Biochemistry Department of Chemistry and Pharmacy, University of Muenster, Münster, Germany

    Henning D. Mootz

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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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  • DOI: https://doi.org/10.1007/978-1-4939-6451-2_9

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