Quantitating the statistical distribution of deuterium incorporation to extend the utility of H/D exchange MS data

Anal Chem. 2006 Jan 1;78(1):207-14. doi: 10.1021/ac050988l.

Abstract

Measuring the statistical distribution of deuterium incorporated into enzymatically derived peptide fragments provides a valuable dimension to hydrogen/deuterium exchange mass spectrometry data. In this paper, we will discuss our improvement to the linear least-squares method for determining this distribution, through the addition of "zeroes" to the end of the deuterated isotopic envelope, to partially compensate for data truncation due to finite instrumental signal-to-noise ratios. The value of the distribution is demonstrated in a simple experimental example, where the linearity between average deuteration and percent D2O used to label test peptides hides a more complex relationship between the site-labeling probability and the total number of sites. This method offers the opportunity to resolve cases where a single peptide experiences distinct, independent biochemical states with each bearing a unique average deuteration; this can occur when a protein is modified to substoichiometric levels. From the experimentally determined distribution of a heterogeneously deuterated peptide, it was possible to extract the average deuteration of each component of the mixture.

MeSH terms

  • Deuterium / chemistry*
  • Deuterium Exchange Measurement / methods*
  • Entropy
  • Hydrogen / chemistry*
  • Peptide Fragments / chemistry
  • Proteins / chemistry*
  • Protons
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*
  • Statistical Distributions

Substances

  • Peptide Fragments
  • Proteins
  • Protons
  • Hydrogen
  • Deuterium