Measurement of backbone hydrogen-deuterium exchange in the type III secretion system needle protein PrgI by solid-state NMR

J Magn Reson. 2017 Oct:283:110-116. doi: 10.1016/j.jmr.2017.08.012. Epub 2017 Sep 1.

Abstract

In this report we present site-specific measurements of amide hydrogen-deuterium exchange rates in a protein in the solid state phase by MAS NMR. Employing perdeuteration, proton detection and a high external magnetic field we could adopt the highly efficient Relax-EXSY protocol previously developed for liquid state NMR. According to this method, we measured the contribution of hydrogen exchange on apparent 15N longitudinal relaxation rates in samples with differing D2O buffer content. Differences in the apparent T1 times allowed us to derive exchange rates for multiple residues in the type III secretion system needle protein.

Keywords: Amide protons; Deuteration; H/D exchange; Magic-angle spinning solid-state NMR; PrgI; Proton detection; Relax-EXSY; Relaxation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry
  • Deuterium / chemistry*
  • Deuterium Exchange Measurement / methods*
  • Deuterium Oxide
  • Electromagnetic Fields
  • Hydrogen / chemistry*
  • Ion Exchange
  • Magnetic Resonance Spectroscopy / methods*
  • Models, Molecular
  • Nitrogen Isotopes
  • Protons

Substances

  • Amides
  • Nitrogen Isotopes
  • Protons
  • Hydrogen
  • Deuterium
  • Deuterium Oxide