Metabolomics

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Isotopomer analysis of MCF-7 breast cancer cell line incubated with 13C labelled glucose (NMR assay)


ABSTRACT: MS and NMR are two of the most powerful spectroscopic techniques used in the field of fluxomics, to analyze the flux distributions in cellular metabolomic pathways. Both of them possess pros and cons, such as, MS being the most sensitive and low-cost technique, while needs extensive sample processing and providing incomplete labeling data, whereas NMR needs lesser sample preparation and provides more detailed labeling data, while being less sensitive and expensive. Since both techniques could be viewed as complementary to each other, combining their data could yield a detailed and complete information about a system under study. In this case, samples were generated from a single experiment and the same set of samples was used both in MS and 1D NMR to analyze the 13C label distributions in TCA cycle intermediates. The data generated from both methods were used with a simulation script that was developed to integrate the MS and NMR data. MCF-7 breast cancer cell line was used as an experimental model and it was incubated with 100% [1,2-13C]-glucose and 100% [U-13C]-glucose for 8 hours. The cells, before and after incubation with tracers, were frozen in liquid nitrogen and then later processed for extracting the TCA cycle intermediates. Same set of samples were separated for GC-MS and NMR based analysis and processed for the respective techniques. The isotopologue analysis was performed by GC-MS and the isotopomer analysis was performed by NMR and both sets of data were integrated with the simulation software. In this study there is the data for NMR analysis. This study contains the data for NMR analysis.

The GC-MS assay for this study can be found in the MetaboLights study MTBLS182.

Linked Studies: MTBLS182

INSTRUMENT(S): Bruker

PROVIDER: MTBLS241 | MetaboLights | 2018-05-02

REPOSITORIES: MetaboLights

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Publications

Combined Analysis of NMR and MS Spectra (CANMS).

Chong Mei M   Jayaraman Anusha A   Marin Silvia S   Selivanov Vitaly V   de Atauri Carulla Pedro R PR   Tennant Daniel A DA   Cascante Marta M   Günther Ulrich L UL   Ludwig Christian C  

Angewandte Chemie (International ed. in English) 20170308 15


Cellular metabolism in mammalian cells represents a challenge for analytical chemistry in the context of current biomedical research. Mass spectrometry and NMR spectroscopy together with computational tools have been used to study metabolism in cells. Compartmentalization of metabolism complicates the interpretation of stable isotope patterns in mammalian cells owing to the superimposition of different pathways contributing to the same pool of analytes. This indicates a need for a model-free app  ...[more]

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