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Extended Gas-Phase Trapping Followed by Surface-Induced Dissociation of Noncovalent Protein Complexes

Author:
Harvey, Sophie R., Yan, Jing, Brown, Jeffery M., Hoyes, Emmy, Wysocki, Vicki H.
Source:
Analytical chemistry 2016 v.88 no.2 pp. 1218-1221
ISSN:
1520-6882
Subject:
C-reactive protein, beta-lactoglobulin, concanavalin A, dissociation, gases, mass spectrometry, phosphopyruvate hydratase, pyruvate kinase, solvents, streptavidin, trapping
Abstract:
Mass spectrometry has emerged as a useful tool in the study of proteins and protein complexes. It is of fundamental interest to explore how the structures of proteins and protein complexes are affected by the absence of solvent and how this alters with increasing time in the gas phase. Here we demonstrate that a range of protein and protein complexes can be confined within the Trap T-wave region of a modified Waters Synapt G2S instrument, including monomeric (β-lactoglobulin), dimeric (β-lactoglobulin and enolase), tetrameric (streptavidin, concanavalin A, and pyruvate kinase), and pentameric (C-reactive protein) complexes, ranging in size up to 237 kDa. We demonstrate that complexes can be confined within the Trap region for varying lengths of time over the range 1–60 s and with up to 86% trapping efficiency for 1 s trapping. Furthermore, using model systems, we show that these noncovalent complexes can also be fragmented by surface-induced dissociation (SID) following trapping. SID reveals similar dissociation patterns over all trapping times studied for unactivated protein complexes, suggesting that any conformational changes occurring over this time scale are insufficient to cause substantial differences in the SID spectra of these complexes. Intentional alteration of structure by cone activation produces a distinct SID spectrum, with the differences observed being conserved, in comparison to unactivated complex, after trapping. However, subtle differences in the SID spectra of the activated complex are also observed as a function of trapping time.
Agid:
5304296