Segmental isotopic labeling of proteins for NMR study using intein technology

Dongsheng Liu, David Cowburn

Research output: Chapter in Book/Report/Conference proceedingChapter

7 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages131-145
Number of pages15
Volume1495
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1495
ISSN (Print)10643745

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Keywords

  • Expressed protein ligation
  • Intein
  • Isotopic labeling
  • NMR
  • Structural biology

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Liu, D., & Cowburn, D. (2017). Segmental isotopic labeling of proteins for NMR study using intein technology. In Methods in Molecular Biology (Vol. 1495, pp. 131-145). (Methods in Molecular Biology; Vol. 1495). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6451-2_9