NR AVJD
AU Nazabal,A.; Schmitter,J.M.
TI Hydrogen-deuterium exchange analyzed by matrix-assisted laser desorption-ionization mass spectrometry and the HET-s prion model
QU Methods in Enzymology 2006; 413: 167-81
PT journal article
AB Hydrogen/deuterium (H/D) exchange analyzed by mass spectrometry (HXMS) is a valuable tool for the investigation of protein conformation and dynamics. After exchange, the sample is generally submitted to electrospray ionization for mass analysis. Matrix-assisted laser desorption ionization (MALDI) has been used in a limited number of studies but has several significant advantages that include simplification of the spectra attributable to a predominance of singly charged ions, speed of analysis, sensitivity, and low H/D back-exchange level. MALDI-HXMS has been used to study amyloid aggregates from the HET-s prion protein. Our results underline the ability of this method to determine solvent accessibility within the amyloid aggregates, reaching a resolution of one to four amino acids. To achieve a complete peptide mass fingerprint of the protein, we have taken benefits of an ion trap operating in liquid chromatography-MS/MS mode. MALDI time-of-flight-MS was then used to determine deuterium incorporation within each peptide along the sequence of HET-s. The combined advantages of these two instruments yield a suitable solution for HXMS experiments that require highly resolved peptide mass fingerprints, high sensitivity, and speed of analysis for deuterium incorporation measurements.
MH Deuterium Exchange Measurement/*methods; Fungal Proteins/*chemistry; Pepsin A/metabolism; Peptide Fragments/chemistry; Peptide Mapping/methods; Prions/*chemistry; Research Support, Non-U.S. Gov't; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/*methods
AD Swiss Federal Institute of Technology, ETH, Department of Chemistry and Applied Biosciences, Zürich, Switzerland.
SP englisch
PO USA