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Analysis of sulfated peptides from the skin secretion of the Pachymedusa dacnicolor frog using IMAC ‐Ga enrichment and high‐resolution mass spectrometry
Author(s) -
Balderrama Griselda Demesa,
Meneses Erika P.,
Orihuela Lorena Hernández,
Hernández Oscar Villa,
Franco Ruben Castro,
Robles Victoria Pando,
Batista Cesar Vicente Ferreira
Publication year - 2011
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.4950
Subject(s) - chemistry , mass spectrometry , sulfation , chromatography , electrospray ionization , tandem mass spectrometry , orbitrap , fragmentation (computing) , dissociation (chemistry) , peptide , matrix assisted laser desorption/ionization , collision induced dissociation , desorption , biochemistry , adsorption , organic chemistry , computer science , operating system
Immobilized metal ion affinity chromatography (IMAC) has been widely used for the enrichment of phosphopeptides, whereas no report exists describing the use of IMAC columns for the enrichment of sulfopeptides. In this study, we used IMAC‐Ga microcolumns for the enrichment of sulfopeptides from a complex mixture of peptides, extracted from skin secretions of the Pachymedusa dacnicolor frog. The enriched fraction obtained by IMAC‐Ga was analyzed by liquid chromatograpy/electrospray ionization mass spectrometry (LC/ESI‐MS) in an Orbitrap XL and by matrix‐assisted laser desorption/ionization time‐of‐flight time‐of‐flight (MALDI‐TOF/TOF) in an ABI 4800 instrument. From this fraction, different sulfated and non‐sulfated peptides belonging to the caerulin and bradykinin families were structurally characterized. Other interesting negatively charged groups, such as phosphate adducts of dermaseptins and pyridoxal phosphate attached to a protease inhibitor, were also characterized. Unexpectedly, some dermaseptin antimicrobial peptides were also enriched by IMAC‐Ga and a Sauvatine‐like peptide was also fully sequenced. Furthermore, neutral loss of sulfated peptides and their fragmentation patterns in the gas phase were also compared using collision‐induced dissociation (CID) and high‐energy collision dissociation (HCD). Our present study provides evidence that IMAC‐Ga enrichment is a fast, useful and promising method for high‐throughput analysis of sulfated‐peptides, since high‐resolution mass spectrometers can be used for this purpose. Copyright © 2011 John Wiley & Sons, Ltd.