2015
DOI: 10.1016/j.ijms.2015.07.008
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Enhancing protein disulfide bond cleavage by UV excitation and electron capture dissociation for top-down mass spectrometry

Abstract: The application of ion pre-activation with 266 nm ultraviolet (UV) laser irradiation combined with electron capture dissociation (ECD) is demonstrated to enhance top-down mass spectrometry sequence coverage of disulfide bond containing proteins. UV-based activation can homolytically cleave a disulfide bond to yield two separated thiol radicals. Activated ECD experiments of insulin and ribonuclease A containing three and four disulfide bonds, respectively, were performed. UV-activation in combination with ECD a… Show more

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Cited by 45 publications
(71 citation statements)
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“…The backbone cleavages preferentially occur near the C-terminus of chain B—which is consistent with the ECD and ETD work of others [8, 10, 14, 24]—and in regions close to aromatic residues, like tyrosine, which also has been noted by others [26]. Compared to ETD, CTD does not produce as many fragment ions near the N-terminus of chain B for 6+ insulin precursor.…”
Section: Resultssupporting
confidence: 89%
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“…The backbone cleavages preferentially occur near the C-terminus of chain B—which is consistent with the ECD and ETD work of others [8, 10, 14, 24]—and in regions close to aromatic residues, like tyrosine, which also has been noted by others [26]. Compared to ETD, CTD does not produce as many fragment ions near the N-terminus of chain B for 6+ insulin precursor.…”
Section: Resultssupporting
confidence: 89%
“…B y 10 2+ , B y 11 2+ , B y 12 2+ , B y 13 2+ and B y 14 2+ (blue font), was also observed from the 6+ precursor, which are rarely observed in other MS/MS experiments. Fragmentation in these region appears rather unique to CTD, since most techniques (CID [7, 8], ETD [14], ECD/AI ECD [24]) struggle to induce fragmentation in this region. The B y 10 2+ and B y 11 2+ fragments are be formed from the cleavage of C-terminus of chain B, outside the inter-chain cyclic region, but the formation of B y 12 2+ , B y 13 2+ and B y 14 2+ requires breakage of both inter-chain disulfide linkages in addition to the backbone cleavage.…”
Section: Resultsmentioning
confidence: 99%
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