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Top-Down Hydrogen–Deuterium Exchange Analysis of Protein Structures Using Ultraviolet Photodissociation
- Source :
- Analytical Chemistry. 90:3079-3082
- Publication Year :
- 2018
- Publisher :
- American Chemical Society (ACS), 2018.
-
Abstract
- Top-down hydrogen-deuterium exchange (HDX) analysis using electron capture or transfer dissociation Fourier transform mass spectrometry (FTMS) is a powerful method for the analysis of secondary structure of proteins in solution. The resolution of the method is a function of the degree of fragmentation of backbone bonds in the proteins. While fragmentation is usually extensive near the N- and C-termini, electron capture (ECD) or electron transfer dissociation (ETD) fragmentation methods sometimes lack good coverage of certain regions of the protein, most often in the middle of the sequence. Ultraviolet photodissociation (UVPD) is a recently developed fast-fragmentation technique, which provides extensive backbone fragmentation that can be complementary in sequence coverage to the aforementioned electron-based fragmentation techniques. Here, we explore the application of electrospray ionization (ESI)-UVPD FTMS on an Orbitrap Fusion Lumos Tribrid mass spectrometer to top-down HDX analysis of proteins. We have incorporated UVPD-specific fragment-ion types and fragment-ion mixtures into our isotopic envelope fitting software (HDX Match) for the top-down HDX analysis. We have shown that UVPD data is complementary to ETD, thus improving the overall resolution when used as a combined approach.
- Subjects :
- Electron capture
Chemistry
Electrospray ionization
010401 analytical chemistry
010402 general chemistry
Photochemistry
Orbitrap
Mass spectrometry
01 natural sciences
Fourier transform ion cyclotron resonance
0104 chemical sciences
Analytical Chemistry
law.invention
Electron-transfer dissociation
Fragmentation (mass spectrometry)
law
Hydrogen–deuterium exchange
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 90
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....635776ed1a4ce07cb2792243f6db8dfb