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A new strategy utilizing electrospray ionization‐quadrupole ion trap mass spectrometry for the qualitative determination of GnRH peptides
Author(s) -
Myers Tanya R.,
Patonay Gabor
Publication year - 2006
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/jms.1054
Subject(s) - chemistry , fragmentation (computing) , electrospray ionization , mass spectrometry , tandem mass spectrometry , peptide , electrospray , ion trap , pentapeptide repeat , chromatography , computational biology , biochemistry , biology , ecology
Numerous forms of the neurotransmitter GnRH have been discovered in vertebrates and invertebrates. Methods used for identification of these peptides are laborious and often require the application of multiple, confirmatory techniques. In this study, we investigate whether HPLC‐MS/MS and de novo sequencing techniques applied to whole peptide analysis can provide a simpler approach to GnRH characterization. Experiments were performed with six GnRH forms (chicken I, chicken II, lamprey III, mammalian, salmon and seabream) to determine whether MS/MS spectra would be dominated by proline‐directed fragmentation to the detriment of obtaining sufficient fragmentation for sequencing. While the expected b 8 fragment was prominent, sufficient ion series were obtained for the six GnRH peptides to provide sequence identification. On the basis of the patterns observed for six model peptides, similar fragmentation patterns are expected for other GnRH forms. To confirm the applicability of the method, extracts from Sprague–Dawley rat brains were examined. These experiments confirm the presence of mammalian GnRH and a posttranslationally modified form of mammalian GnRH, hydroxyproline 9 GnRH, in Sprague–Dawley rat brains and demonstrate that ESI‐MS/MS techniques provide a valuable addition to existing qualitative methods. Copyright © 2006 John Wiley & Sons, Ltd.