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13C labelling of nematode worms to improve metabolome coverage by heteronuclear nuclear magnetic resonance experiments

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Title: 13C labelling of nematode worms to improve metabolome coverage by heteronuclear nuclear magnetic resonance experiments
Authors: Geier, F
Leroi, A
Bundy, J
Item Type: Journal Article
Abstract: Nuclear magnetic resonance (NMR) spectroscopy is widely used as a metabolomics tool, and 1D spectroscopy is overwhelmingly the commonest approach. The use of 2D spectroscopy could offer significant advantages in terms of increased spectral dispersion of peaks, but has a number of disadvantages—in particular, heteronuclear 2D spectroscopy is often much less sensitive than 1D NMR. One factor contributing to this low sensitivity in 13C/1H heteronuclear NMR is the low natural abundance of the 13C stable isotope; as a consequence, where it is possible to label biological material with 13C, there is a potential enhancement of sensitivity of up to around 90fold. However, there are some problems that can reduce the advantages otherwise gained—in particular, the fine structure arising from 13C/13C coupling, which is essentially non-existent at natural abundance, can reduce the possible sensitivity gain and increase the chances of peak overlap. Here, we examined the use of two different heteronuclear single quantum coherence (HSQC) pulse sequences for the analysis of fully 13C-labeled tissue extracts from Caenorhabditis elegans nematodes. The constant time ct-HSQC had improved peak shape, and consequent better peak detection of metabolites from a labeled extract; matching this against reference spectra from the HMDB gave a match to about 300 records (although fewer actual metabolites, as some of these represent false positive matches). This approach gives a rapid and automated initial metabolome assignment, forming an ideal basis for further manual curation.
Issue Date: 26-Apr-2019
Date of Acceptance: 4-Apr-2019
URI: http://hdl.handle.net/10044/1/70080
DOI: https://dx.doi.org/10.3389/fmolb.2019.00027
ISSN: 2296-889X
Publisher: Frontiers Media
Journal / Book Title: Frontiers in Molecular Biosciences
Volume: 6
Copyright Statement: © 2019 Geier, Leroi and Bundy. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Sponsor/Funder: AXA SA- AXA Research Fund
Funder's Grant Number: 200-Doc-Imperial College-Geier
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
C. elegans
HSQC
CT-HSQC
sensitivity
metabolomics
metabolome
NMR metabolomics
labeling
CAENORHABDITIS-ELEGANS
COMPLEX-MIXTURES
NMR-SPECTRA
2D NMR
HMDB
DATABASE
UNCOVERS
C. elegans
CT-HSQC
HSQC
NMR metabolomics
labeling
metabolome
metabolomics
sensitivity
Publication Status: Published
Article Number: ARTN 26
Appears in Collections:Division of Surgery



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