Isotope labeling in insect cells

Methods Mol Biol. 2012:831:37-54. doi: 10.1007/978-1-61779-480-3_3.

Abstract

Recent years have seen remarkable progress in applying nuclear magnetic resonance (NMR) spectroscopy to proteins that have traditionally been difficult to study due to issues with folding, posttranslational modification, and expression levels or combinations thereof. In particular, insect cells have proved useful in allowing large quantities of isotope-labeled, functional proteins to be obtained and purified to homogeneity, allowing study of their structures and dynamics by using NMR. Here, we provide protocols that have proven successful in such endeavors.

MeSH terms

  • Amino Acids / metabolism
  • Animals
  • Baculoviridae
  • Bioreactors*
  • Cell Line
  • Genetic Vectors / genetics
  • Molecular Chaperones / metabolism
  • Nitrogen Isotopes / metabolism
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Protein Folding
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism*
  • Spodoptera
  • Temperature
  • Transfection / methods

Substances

  • Amino Acids
  • Molecular Chaperones
  • Nitrogen Isotopes
  • Recombinant Proteins