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• W2071139100 abstract "Characterization of protein cross-linking, particularly without prior knowledge of the chemical nature and site of cross-linking, poses a significant challenge, because of their intrinsic structural complexity and the lack of a comprehensive analytical approach. Toward this end, we have developed a generally applicable workflow—XChem-Finder—that involves four stages: (1) detection of cross-linked peptides via 18O-labeling at C-termini; (2) determination of the putative partial sequences of each cross-linked peptide pair using a fragment ion mass database search against known protein sequences coupled with a de novo sequence tag search; (3) extension to full sequences based on protease specificity, the unique combination of mass, and other constraints; and (4) deduction of cross-linking chemistry and site. The mass difference between the sum of two putative full-length peptides and the cross-linked peptide provides the formulas (elemental composition analysis) for the functional groups involved in each cross-linking. Combined with sequence restraint from MS/MS data, plausible cross-linking chemistry and site were inferred, and ultimately confirmed, by matching with all data. Applying our approach to a stressed IgG2 antibody, 10 cross-linked peptides were discovered and found to be connected via thioethers originating from disulfides at locations that had not been previously recognized. Furthermore, once the cross-link chemistry was revealed, a targeted cross-link search yielded 4 additional cross-linked peptides that all contain the C-terminus of the light chain." @default.
• W2071139100 created "2016-06-24" @default.
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• W2071139100 date "2013-05-28" @default.
• W2071139100 modified "2023-10-16" @default.
• W2071139100 title "Discovery of Undefined Protein Cross-Linking Chemistry: A Comprehensive Methodology Utilizing ¹⁸O-Labeling and Mass Spectrometry" @default.
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• W2071139100 doi "https://doi.org/10.1021/ac400666p" @default.
• W2071139100 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3691076" @default.
• W2071139100 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/23634697" @default.
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