Synthesis of multi-unit protein hetero-complexes in the gas phase via ion-ion chemistry

Journal of Mass Spectrometry : JMS
Harsha P Gunawardena, Scott A McLuckey

Abstract

The synthesis of protein hetero-complex ions via ion-ion reactions in the gas phase is demonstrated in a quadrupole ion trap. Bovine cytochrome c cations and bovine ubiquitin anions are used as reactant species in the stepwise construction of complexes containing as many as six protein sub-units. For any set of reactants, a series of competitive and consecutive reactions is possible. The yield of complex ions for any given sequence of reactions is primarily limited by the presence of competitive reactions. Proton transfer represents the most important competitive reaction that adversely affects protein complex synthesis. In the present data, proton transfer takes place most extensively in the first step of complex synthesis, when single protein sub-units are subjected to reaction with one another. Proton transfer is found to be less extensive when one of the reactants is a protein complex. The generation of hexameric hetero-complexes containing two cytochrome c molecules and four ubiquitin molecules is demonstrated with two different synthesis approaches. The first involved the initial reaction of several charge states of cytochrome c and several charges states of ubiquitin. The sequence of reactions in this example illustrates...Continue Reading

Citations

Sep 27, 2005·Journal of the American Society for Mass Spectrometry·Yu XiaScott A McLuckey
Mar 14, 2007·Journal of the American Society for Mass Spectrometry·Xiaorong Liang, Scott A McLuckey
Dec 18, 2007·Journal of the American Society for Mass Spectrometry·Yu Xia, Scott A McLuckey
Jul 20, 2010·Annual Review of Analytical Chemistry·Teng-Yi Huang, Scott A McLuckey
Dec 22, 2012·Chemical Communications : Chem Comm·Boone M Prentice, Scott A McLuckey
Jun 1, 2005·Analytical Chemistry·Yu XiaScott A McLuckey
May 19, 2020·Journal of the American Society for Mass Spectrometry·Hsi-Chun ChaoScott A McLuckey

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