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Electron Detachment Dissociation for Top‐Down Mass Spectrometry of Acidic Proteins
Author(s) -
Ganisl Barbara,
Valovka Taras,
Hartl Markus,
Taucher Monika,
Bister Klaus,
Breuker Kathrin
Publication year - 2011
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201003709
Subject(s) - electron capture dissociation , chemistry , cysteine , mass spectrometry , side chain , dissociation (chemistry) , electron transfer dissociation , fragmentation (computing) , tandem mass spectrometry , limiting , lysozyme , amino acid , oligonucleotide , chromatography , combinatorial chemistry , biochemistry , organic chemistry , enzyme , dna , biology , mechanical engineering , ecology , engineering , polymer
Electron detachment dissociation (EDD) is an emerging mass spectrometry (MS) technique for the primary structure analysis of peptides, carbohydrates, and oligonucleotides. Herein, we explore the potential of EDD for sequencing of proteins of up to 147 amino acid residues by using top‐down MS. Sequence coverage ranged from 72 % for Melittin, which lacks carboxylic acid functionalities, to 19 % for an acidic 147‐residue protein, to 12 % for Ferredoxin, which showed unusual backbone fragmentation next to cysteine residues. A limiting factor for protein sequencing by EDD is the facile loss of small molecules from amino acid side chains, in particular CO 2 . Based on the types of fragments observed and fragmentation patterns found, we propose detailed mechanisms for protein backbone cleavage and side chain dissociation in EDD. The insights from this study should further the development of EDD for top‐down MS of acidic proteins.