Journal article
Hydrogen-deuterium exchange analyzed by matrix-assisted laser desorption-ionization mass spectrometry and the HET-s prion model.
- Nazabal A Swiss Federal Institute of Technology, ETH, Department of Chemistry and Applied Biosciences, Zurich, Switzerland.
- Schmitter JM
- 2006-10-19
Published in:
- Methods in enzymology. - 2006
Deuterium Exchange Measurement
Fungal Proteins
Pepsin A
Peptide Fragments
Peptide Mapping
Prions
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
English
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.
- Language
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- English
- Open access status
- closed
- Identifiers
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- DOI 10.1016/S0076-6879(06)13009-8
- PMID 17046396
- Persistent URL
- https://sonar.ch/global/documents/123598
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