Measuring the Hydrogen/Deuterium Exchange of Proteins at High Spatial Resolution by Mass Spectrometry: Overcoming Gas-Phase Hydrogen/Deuterium Scrambling

2014 ◽  
Vol 47 (10) ◽  
pp. 3018-3027 ◽  
Author(s):  
Kasper D. Rand ◽  
Martin Zehl ◽  
Thomas J. D. Jørgensen
Keyword(s):  
Mass Spectrometry ◽  
Gas Phase ◽  
Spatial Resolution ◽  
High Spatial Resolution ◽  
Deuterium Exchange ◽  
Hydrogen Deuterium Exchange ◽  
Hydrogen Deuterium

Do electrospray mass spectra reflect the ligand binding state of proteins in solution?

2005 ◽  
Vol 83 (11) ◽  
pp. 1953-1960 ◽  
Author(s):  
Belal M Hossain ◽  
Douglas A Simmons ◽  
Lars Konermann
Keyword(s):  
Mass Spectrometry ◽  
Gas Phase ◽  
Protein Interactions ◽  
Noncovalent Complex ◽  
Deuterium Exchange ◽  
Solution Phase ◽  
Hydrogen Deuterium Exchange ◽  
Esi Ms ◽  
Protein Ions ◽  
Hydrogen Deuterium

Electrospray ionization (ESI) mass spectrometry (MS) has become a popular tool for monitoring ligand–protein and protein–protein interactions. Due to the "gentle" nature of the ionization process, it is often possible to transfer weakly bound complexes into the gas phase, thus making them amenable to MS detection. One problem with this technique is the potential occurrence of fragmentation events during ESI. Also, some analytes tend to cluster together during ionization, thus forming nonspecific gas-phase assemblies that do not represent solution-phase complexes. In this work, we implemented a hydrogen–deuterium exchange (HDX) approach that can reveal whether or not the free and (or) bound constituents of a complex observed in ESI-MS reflect the binding situation in solution. Proteins are subjected to ESI immediately following an isotopic labeling pulse; only ligand-free and ligand-bound protein ions that were formed directly from the corresponding solution-phase species showed different HDX levels. Using myoglobin as a model system, it is demonstrated that this approach can readily distinguish scenarios where the heme–protein interactions were disrupted in solution from those where dissociation of the complex occurred in the gas phase. Experiments on cytochrome c strongly suggest that dimeric protein ions observed in ESI-MS reflect aggregates that were formed in solution.Key words: electrospray mass spectrometry, ligand–protein interaction, noncovalent complex, hydrogen–deuterium exchange, protein folding.

scholarly journals High-Precision, Gas-Phase Hydrogen/Deuterium-Exchange Kinetics by Mass Spectrometry Enabled by Exchange Standards

2020 ◽  
Vol 92 (11) ◽  
pp. 7725-7732 ◽  
Author(s):  
Sanjit S. Uppal ◽  
Abhigya Mookherjee ◽  
Rick Harkewicz ◽  
Sarah E. Beasley ◽  
Matthew F. Bush ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Phase ◽  
High Precision ◽  
Deuterium Exchange ◽  
Hydrogen Deuterium Exchange ◽  
Hydrogen Deuterium ◽  
Exchange Kinetics

Solution and gas phase hydrogen/deuterium exchange of oligoamide complexes by nanoelectrospray mass spectrometry

2007 ◽  
Vol 261 (1) ◽  
pp. 13-24 ◽  
Author(s):  
Honghai Jiang ◽  
Anthony R. Dolan ◽  
Minfeng Li ◽  
Marie A. Moy ◽  
Bing Gong ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Phase ◽  
Deuterium Exchange ◽  
Hydrogen Deuterium Exchange ◽  
Hydrogen Deuterium
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