A triaxial probe for on-line proteolysis coupled with hydrogen/deuterium exchange-electrospray mass spectrometry

J Am Soc Mass Spectrom. 2007 Feb;18(2):208-17. doi: 10.1016/j.jasms.2006.09.011. Epub 2006 Oct 30.

Abstract

An on-line proteolysis system utilizing a triaxial electrospray probe was developed to aid localization of the hydrogen-bonding interaction sites in hydrogen/deuterium exchange-mass spectrometry (HDX-MS) studies of Abeta (1-40) fibrils. The probe allows delayed introduction of the organic solvent component needed for stable electrospray, thus enhancing hydrolysis performance relative to that of a coaxial probe. Effective on-line digestion was accomplished in approximately 12 s. The probe should be of general utility for HDX-MS studies of amyloid fibrils and other protein aggregates.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amyloid beta-Peptides / chemistry*
  • Animals
  • Deuterium Exchange Measurement*
  • Hydrogen / chemistry*
  • Hydrogen Bonding
  • Hydrolysis
  • Pepsin A / chemistry
  • Peptide Fragments / chemistry*
  • Peptide Mapping
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Swine

Substances

  • Amyloid beta-Peptides
  • Peptide Fragments
  • amyloid beta-protein (1-40)
  • Hydrogen
  • Pepsin A