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Sequencing of a branched peptide using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Fournier I.,
Chaurand P.,
Bolbach G.,
Lützenkirchen F.,
Spengler B.,
Tabet J. C.
Publication year - 2000
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/1096-9888(200012)35:12<1425::aid-jms77>3.0.co;2-c
Subject(s) - chemistry , mass spectrometry , peptide , matrix assisted laser desorption/ionization , protein mass spectrometry , mass spectrum , chromatography , peptide sequence , ionization , desorption , time of flight mass spectrometry , acetylation , combinatorial chemistry , analytical chemistry (journal) , tandem mass spectrometry , ion , organic chemistry , biochemistry , adsorption , gene
Chemical degradation methods combined with matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) and post‐source decay (PSD)‐MALDI reflex TOF mass spectrometry (MS) were used to determine the sequence of a peptide branched on to a known peptide backbone. This study was applied to a branched peptide model (derivative of substance P). The branched peptide mimics a digest of a membrane receptor on to which a derivative of substance P was photochemically linked. Chemical degradation based on N ‐terminal ladder sequencing in combination with MALDI‐TOF‐MS gave only partial sequence information. Although single PSD mass spectra still remain difficult to interpret unambiguously, PSD‐MALDI‐TOF‐MS was combined with on‐target acetylation and H – D exchange to give a better and successful approach to the unambiguous determination of the complete amino acid side‐chain sequence. This study shows the capability of MALDI‐TOF‐MS to help in characterizing ligand–receptor interactions. Copyright © 2000 John Wiley & Sons, Ltd.