Methyl-Specific Isotope Labeling Strategies for NMR Studies of Membrane Proteins

Methods Mol Biol. 2017:1635:109-123. doi: 10.1007/978-1-4939-7151-0_6.

Abstract

Methyl groups are very useful probes of structure, dynamics, and interactions in protein NMR spectroscopy. In particular, methyl-directed experiments provide high sensitivity even in very large proteins, such as membrane proteins in a membrane-mimicking environment. In this chapter, we discuss the approach for labeling methyl groups in E. coli-based protein expression, as exemplified with the mitochondrial carrier GGC.

Keywords: Detergent; Deuteration; Methyl labeling; NMR spectroscopy; Refolding.

MeSH terms

  • Carbon Isotopes / chemistry
  • Deuterium / chemistry
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Isotope Labeling / methods*
  • Membrane Proteins / chemistry*
  • Membrane Proteins / metabolism
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Refolding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Carbon Isotopes
  • Membrane Proteins
  • Recombinant Proteins
  • Deuterium