Rapid sensitive analysis of cysteine rich peptide venom components

Ueberheide, B. M., Fenyo, D, Alewood, P. F. and Chait, B. T. (2009) Rapid sensitive analysis of cysteine rich peptide venom components. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 106 17: 6910-6915. doi:10.1073/pnas.0900745106


Author Ueberheide, B. M.
Fenyo, D
Alewood, P. F.
Chait, B. T.
Title Rapid sensitive analysis of cysteine rich peptide venom components
Journal name PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   Check publisher's open access policy
ISSN 0027-8424
Publication date 2009-04-01
Year available 2009
Sub-type Article (original research)
DOI 10.1073/pnas.0900745106
Open Access Status Not Open Access
Volume 106
Issue 17
Start page 6910
End page 6915
Total pages 6
Editor Randy Schekman
Place of publication Washington , D.C., U.S.A.
Publisher National Academy of Sciences
Language eng
Subject C1
970103 Expanding Knowledge in the Chemical Sciences
0304 Medicinal and Biomolecular Chemistry
Abstract Disulfide-rich peptide venoms from animals such as snakes, spiders, scorpions, and certain marine snails represent one of nature's great diversity libraries of bioactive molecules. The various species of marine cone shells have alone been estimated to produce >50,000 distinct peptide venoms. These peptides have stimulated considerable interest because of their ability to potently alter the function of specific ion channels. To date, only a small fraction of this immense resource has been characterized because of the difficulty in elucidating their primary structures, which range in size between 10 and 80 aa, include up to 5 disulfide bonds, and can contain extensive posttranslational modifications. The extraordinary complexity of crude venoms and the lack of DNA databases for many of the organisms of interest present major analytical challenges. Here, we describe a strategy that uses mass spectrometry for the elucidation of the mature peptide toxin components of crude venom samples. Key to this strategy is our use of electron transfer dissociation (ETD), a mass spectrometric fragmentation technique that can produce sequence information across the entire peptide backbone. However, because ETD only yields comprehensive sequence coverage when the charge state of the precursor peptide ion is sufficiently high and the m/z ratio is low, we combined ETD with a targeted chemical derivatization strategy to increase the charge state of cysteine-containing peptide toxins. Using this strategy, we obtained full sequences for 31 peptide toxins, using just 7% of the crude venom from the venom gland of a single cone snail (Conus textile).
Keyword conotoxins
ELECTRON-CAPTURE DISSOCIATION
cysteine derivatization
de novo sequencing
electron transfer dissociation
Mass Spectrometry
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID RR00862
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
Institute for Molecular Bioscience - Publications
 
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Created: Thu, 03 Sep 2009, 18:12:39 EST by Mr Andrew Martlew on behalf of Institute for Molecular Bioscience