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Deducing disulfide patterns of cysteine-rich proteins using signature fragments produced by top-down mass spectrometry

TitleDeducing disulfide patterns of cysteine-rich proteins using signature fragments produced by top-down mass spectrometry
Publication TypeJournal Article
Year of Publication2018
AuthorsZhao, X, Shen, Y, Tong, W, Wang, G, Chen, DDa Yong
JournalAnalyst
Volume143
Start Page817
Pagination817-823
Date Published01/2018
Abstract

Direct mapping of protein disulfide pattern using top-down mass spectrometry (MS) is often hampered by inadequate fragmentation at the disulfide-enclosing region{,} and insufficient structural information provided by the fragments. Here we used electron-transfer/high energy collision dissociation (EThcD) to improve the fragmentation efficiency{,} and developed strategies that minimize false positive identification of fragments and deconvolute signals representing specific modifications made to the disulfide-cleavage-induced fragments. We observed clear correlations between unique modification (attachment or removal of H or SH) pattern and the number of disulfide bonds that enclose the corresponding region. Using the characteristic signature fragments{,} we in part localized Cys-bridging sites in disulfide-scrambled lysozyme{,} and reduced the number of putative disulfide patterns from 104 to 6. The results demonstrated the feasibility of direct analysis of complex disulfide patterns using top-down MS.

URLhttp://dx.doi.org/10.1039/C7AN01625E
DOI10.1039/C7AN01625E