DNP-Assisted NMR Investigation of Proteins at Endogenous Levels in Cellular Milieu

Whitney N. Costello, Yiling Xiao, Kendra K. Frederick

Research output: Contribution to journalArticle

Abstract

Structural investigations of biomolecules are typically confined to in vitro systems under extremely limited conditions. These investigations yield invaluable insights, but such experiments cannot capture important structural features imposed by cellular environments. Structural studies of proteins in their native contexts are not only possible using state-of-the-art sensitivity-enhanced (dynamic nuclear polarization, DNP) solid-state nuclear magnetic resonance (NMR) techniques, but these studies also demonstrate that the cellular context can and does have a dramatic influence on protein structure. In this chapter, we describe methods to prepare samples of isotopically labeled proteins at endogenous levels in cellular contexts alongside quality control methods to ensure that such samples accurately model important features of the cellular environment.

Original languageEnglish (US)
JournalMethods in Enzymology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Polarization
Proteins
Biomolecules
Quality Control
Quality control
Experiments
In Vitro Techniques

Keywords

  • Amyloids
  • Dynamic nuclear polarization
  • In situ structural biology
  • In-cell NMR
  • NMR
  • Prions
  • Sup35

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

DNP-Assisted NMR Investigation of Proteins at Endogenous Levels in Cellular Milieu. / Costello, Whitney N.; Xiao, Yiling; Frederick, Kendra K.

In: Methods in Enzymology, 01.01.2018.

Research output: Contribution to journalArticle

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