Premium
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
Author(s) -
Satake Hiroyuki,
Manri Naomi,
Kaneko Akihito,
Hirabayashi Atsumu,
Hasegawa Hideki,
Hashimoto Yuichiro,
Baba Takashi,
Sakamoto Takeshi,
Masuda Katsuyoshi
Publication year - 2013
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.6731
Subject(s) - chemistry , quadrupole ion trap , ion , electron capture dissociation , dissociation (chemistry) , fragmentation (computing) , electron transfer dissociation , ion trap , mass spectrometry , peptide , analytical chemistry (journal) , tandem mass spectrometry , chromatography , biochemistry , organic chemistry , computer science , operating system
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.