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• W1975587425 abstract "Hydrogen exchange in solution combined with ion fragmentation in the gas phase followed by MS detection emerged in recent years as a powerful tool to study higher order protein structure and dynamics. However, a certain type of ion chemistry in the gas phase, namely, internal rearrangement of labile hydrogen atoms (the so-called hydrogen scrambling), is often cited as a factor limiting the utility of this experimental technique. Although several studies have been carried out to elucidate the roles played by various factors in the occurrence and the extent of hydrogen scrambling, there is still no consensus as to what experimental protocol should be followed to avoid or minimize it. In this study we employ fragmentation of mass-selected subpopulations of protein ions to assess the extent of internal proton mobility prior to dissociation. A unique advantage of tandem MS is that it not only provides a means to map the deuterium content of protein ions whose overall levels of isotope incorporation can be precisely defined by controlling the mass selection window, but also correlates this spatial isotope distribution with such global characteristic as the protein ion charge state. Hydrogen scrambling does not occur when the charge state of the precursor protein ions selected for fragmentation is high. Fragment ions derived from both N- and C-terminal parts of the protein are equally unaffected by scrambling. However, spatial distribution of deuterium atoms obtained by fragmenting low-charge-density protein ions is consistent with a very high degree of scrambling prior to the dissociation events. The extent of hydrogen scrambling is also high when multistage fragmentation is used to probe deuterium incorporation locally. Taken together, the experimental results provide a coherent picture of intramolecular processes occurring prior to the dissociation event and provide guidance for the design of experiments whose outcome is unaffected by hydrogen scrambling." @default.
• W1975587425 created "2016-06-24" @default.
• W1975587425 creator A5070800706 @default.
• W1975587425 creator A5077165139 @default.
• W1975587425 date "2010-01-07" @default.
• W1975587425 modified "2023-10-13" @default.
• W1975587425 title "Controlling Hydrogen Scrambling in Multiply Charged Protein Ions during Collisional Activation: Implications for Top-Down Hydrogen/Deuterium Exchange MS Utilizing Collisional Activation in the Gas Phase" @default.
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• W1975587425 doi "https://doi.org/10.1021/ac9021874" @default.
• W1975587425 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2813994" @default.
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