Nuclear magnetic resonance seq (NMRseq): a new approach to peptide sequence tags

Wilson, David, and Daly, Norelle L. (2018) Nuclear magnetic resonance seq (NMRseq): a new approach to peptide sequence tags. Toxins, 10 (11). 437. pp. 1-10.

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Abstract

Structural analysis of peptides with nuclear magnetic resonance (NMR) spectroscopy generally relies on knowledge of the primary sequence to enable assignment of the resonances prior to determination of the three-dimensional structure. Resonance assignment without knowledge of the sequence is complicated by redundancy in amino acid type, making complete de novo sequencing using NMR spectroscopy unlikely to be feasible. Despite this redundancy, we show here that NMR spectroscopy can be used to identify short sequence tags that can be used to elucidate full-length peptide sequences via database searching. In the current study, we have used this approach to identify conotoxins from the venom of the cone snail Conus geographus and determined the three-dimensional structure of a member of the I3 superfamily. This approach is most likely to be useful for the characterization of disulfide-rich peptides, such as those that were chosen for this study, as they generally have well-defined structures, which enhances the quality of the NMR spectra. In contrast to other sequencing methods, the lack of sample manipulation, such as protease digestion, allows for subsequent bioassays to be carried out using the native sample used for sequence identification.

Item ID: 56586
Item Type: Article (Research - C1)
ISSN: 2072-6651
Keywords: NMR spectroscopy, disulfide-rich peptide, conotoxin
Copyright Information: © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Funders: James Cook University (JCU), Australian Research Council (ARC)
Projects and Grants: JCU Capacity Building Grant, ARC LE120100015, ARC LE160100218
Date Deposited: 12 Dec 2018 07:38
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1101 Medical Biochemistry and Metabolomics > 110106 Medical Biochemistry: Proteins and Peptides (incl Medical Proteomics) @ 100%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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