Elucidating the sequence of intact bioactive peptides by using electron capture dissociation and hot electron capture dissociation in a linear radio‐frequency quadrupole ion trap. (15th December 2013)

• Record Type: Journal Article
• Authors: Satake, Hiroyuki; Manri, Naomi; Kaneko, Akihito; Hirabayashi, Atsumu; Hasegawa, Hideki; Hashimoto, Yuichiro; Baba, Takashi; Sakamoto, Takeshi; Masuda, Katsuyoshi
• Is Part Of: Rapid communications in mass spectrometry. Volume 27:Number 23(2013)
• Page Start: 2710
• Page End: 2716
• Publication Date: 2013-12-15
• Subjects: Mass spectrometry -- Periodicals; 543.65
• DOI: 10.1002/rcm.6731
• Languages: English
• ISSNs: 0951-4198

Abstract

Abstract : RATIONALE: Electron capture dissociation (ECD) is useful tool for sequencing of peptides and proteins with post‐translational modifications. To increase the sequence coverage for peptides and proteins, it is important to develop ECD device with high fragmentation efficiency. METHODS: Sequence analysis of intact undigested bioactive peptides (3000–5000 Da) was performed by use of electron capture dissociation (rf‐ECD) and collision‐induced dissociation (CID) in a linear radio‐frequency quadrupole ion trap that was coupled to a time‐of‐flight mass spectrometer. We applied rf‐ECD, hot rf‐ECD (rf‐ECD with high electron energy), and CID for intact bioactive peptide ions of various charge states and evaluated the sequence coverage of their fragment spectra. RESULTS: Hot rf‐ECD produced a higher number of c‐ and z‐type fragment ions of modified peptide ions as electron energy increased in lower charged peptide ions, and sequence coverage greater than 80% was obtained compared with the CID case (40–80%). CONCLUSIONS: The result indicates that intact bioactive modified peptides (Ghrelin, ANP) were correctly identified by use of hot rf‐ECD. Copyright © 2013 John Wiley & Sons, Ltd.