Pulsed hydrogen/deuterium exchange MS/MS for studying the relationship between noncovalent protein complexes in solution and in the gas phase after electrospray ionization

Analytical Chemistry
Belal M Hossain, Lars Konermann

Abstract

Electrospray ionization mass spectrometry (ESI-MS) has become a standard method for monitoring noncovalent protein-protein interactions. Studies employing this approach tend to operate on the premise that the ionic species observed in the mass spectrum directly reflect the corresponding solution-phase protein quaternary structures. However, dissociation or clustering events taking place during ESI may lead to disparities between the ions observed in the mass spectrum and the protein binding state in bulk solution. Recognizing the occurrence of dissociation or clustering artifacts is not straightforward, leading to possible ambiguities in the interpretation of ESI-MS data. This work employs on-line pulsed hydrogen-deuterium exchange (HDX) for probing the origin of various species in the ESI mass spectrum of hemoglobin. In addition to the canonical hemoglobin tetramer, ESI-MS reveals the presence of monomers, dimers, hexamers, and octamers. Tandem mass spectrometry (MS/MS) is used for extracting HDX levels in a subunit-specific manner. Dimeric species exhibit exchange levels that are significantly above those of the tetramer. Monomeric hemoglobin subunits are labeled to an even greater extent. This HDX pattern implies that monome...Continue Reading

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Citations

Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Yang KangD J Douglas
Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Jiangjiang Liu, Lars Konermann
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Yang Kang, D J Douglas
Oct 19, 2011·Journal of the American Society for Mass Spectrometry·Yang KangD J Douglas
Jan 25, 2012·Journal of the American Society for Mass Spectrometry·Elena N KitovaJohn S Klassen
Apr 19, 2013·Journal of the American Society for Mass Spectrometry·Khadijeh Rajabi, D J Douglas
Sep 19, 2006·Journal of the American Society for Mass Spectrometry·Brian L Boys, Lars Konermann
Nov 11, 2008·Journal of the American Society for Mass Spectrometry·John P HodkinsonAlison E Ashcroft
Dec 23, 2008·Journal of the American Society for Mass Spectrometry·P John Wright, D J Douglas
Dec 27, 2008·Journal of the American Society for Mass Spectrometry·Charlotte A ScarffJames H Scrivens
Feb 10, 2012·Journal of the Royal Society, Interface·Gillian R Hilton, Justin L P Benesch
Aug 13, 2009·Mass Spectrometry Reviews·Lars KonermannXin Tong
Dec 1, 2006·Journal of Mass Spectrometry : JMS
Oct 31, 2020·Journal of the American Society for Mass Spectrometry·Alexander I M SeverLars Konermann
Jul 26, 2007·Chemical Reviews·Justin L P BeneschCarol V Robinson

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