Assessing the potential of quantitative 2D HSQC NMR in 13C enriched living organisms View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-02

AUTHORS

Daniel Lane, Thomas E. Skinner, Naum I. Gershenzon, Wolfgang Bermel, Ronald Soong, Rudraksha Dutta Majumdar, Yalda Liaghati Mobarhan, Sebastian Schmidt, Hermann Heumann, Martine Monette, Myrna J. Simpson, André J. Simpson

ABSTRACT

In vivo Nuclear Magnetic Resonance (NMR) spectroscopy has great potential to interpret the biochemical response of organisms to their environment, thus making it an essential tool in understanding toxic mechanisms. However, magnetic susceptibility distortions lead to 1D NMR spectra of living organisms with lines that are too broad to identify and quantify metabolites, necessitating the use of 2D 1H-13C Heteronuclear Single Quantum Coherence (HSQC) as a primary tool. While quantitative 2D HSQC is well established, to our knowledge it has yet to be applied in vivo. This study represents a simple pilot study that compares two of the most popular quantitative 2D HSQC approaches to determine if quantitative results can be directly obtained in vivo in isotopically enriched Daphnia magna (water flea). The results show the perfect-HSQC experiment performs very well in vivo, but the decoupling scheme used is critical for accurate quantitation. An improved decoupling approach derived using optimal control theory is presented here that improves the accuracy of metabolite concentrations that can be extracted in vivo down to micromolar concentrations. When combined with 2D Electronic Reference To access In vivo Concentrations (ERETIC) protocols, the protocol allows for the direct extraction of in vivo metabolite concentrations without the use of internal standards that can be detrimental to living organisms. Extracting absolute metabolic concentrations in vivo is an important first step and should, for example, be important for the parameterization as well as the validation of metabolic flux models in the future. More... »

PAGES

31-42

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s10858-018-0221-2

DOI

http://dx.doi.org/10.1007/s10858-018-0221-2

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1111051993

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30600417


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