High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations]
Tracing the fate of stable isotopically-enriched nutrients is a sophisticated method of describing and quantifying the activity of metabolic pathways. Nuclear Magnetic Resonance (NMR) offers high resolution data, yet is under-utilised due to length of time required to collect the data, quantificatio...
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doaj-f7772f58ab35446bb8f62653581dd6c82020-11-24T22:38:53ZengWellcomeWellcome Open Research2398-502X2018-01-01310.12688/wellcomeopenres.13387.114535High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations]Thomas Brendan Smith0Kamlesh Patel1Haydn Munford2Andrew Peet3Daniel A. Tennant4Mark Jeeves5Christian Ludwig6Institute of Metabolism and Systems Research, University of Birmingham, West Midlands, UKInstitute of Metabolism and Systems Research, University of Birmingham, West Midlands, UKInstitute of Metabolism and Systems Research, University of Birmingham, West Midlands, UKInstitute of Cancer and Genomic Sciences, University of Birmingham, West Midlands, UKInstitute of Metabolism and Systems Research, University of Birmingham, West Midlands, UKInstitute of Cancer and Genomic Sciences, University of Birmingham, West Midlands, UKInstitute of Metabolism and Systems Research, University of Birmingham, West Midlands, UKTracing the fate of stable isotopically-enriched nutrients is a sophisticated method of describing and quantifying the activity of metabolic pathways. Nuclear Magnetic Resonance (NMR) offers high resolution data, yet is under-utilised due to length of time required to collect the data, quantification requiring multiple samples and complicated analysis. Here we present two techniques, quantitative spectral filters and enhancement of the splitting due to J-coupling in 1H,13C-HSQC NMR spectra, which allow the rapid collection of NMR data in a quantitative manner on a single sample. The reduced duration of HSQC spectra data acquisition opens up the possibility of real-time tracing of metabolism including the study of metabolic pathways in vivo. We show how these novel techniques can be used to trace the fate of labelled nutrients in a whole organ model of kidney preservation prior to transplantation using a porcine kidney as a model organ, and also show how the use of multiple nutrients, differentially labelled with 13C and 15N, can be used to provide additional information with which to profile metabolic pathways.https://wellcomeopenresearch.org/articles/3-5/v1 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Thomas Brendan Smith Kamlesh Patel Haydn Munford Andrew Peet Daniel A. Tennant Mark Jeeves Christian Ludwig |
spellingShingle |
Thomas Brendan Smith Kamlesh Patel Haydn Munford Andrew Peet Daniel A. Tennant Mark Jeeves Christian Ludwig High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] Wellcome Open Research |
author_facet |
Thomas Brendan Smith Kamlesh Patel Haydn Munford Andrew Peet Daniel A. Tennant Mark Jeeves Christian Ludwig |
author_sort |
Thomas Brendan Smith |
title |
High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] |
title_short |
High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] |
title_full |
High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] |
title_fullStr |
High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] |
title_full_unstemmed |
High-Speed Tracer Analysis of Metabolism (HS-TrAM) [version 1; referees: 1 approved, 2 approved with reservations] |
title_sort |
high-speed tracer analysis of metabolism (hs-tram) [version 1; referees: 1 approved, 2 approved with reservations] |
publisher |
Wellcome |
series |
Wellcome Open Research |
issn |
2398-502X |
publishDate |
2018-01-01 |
description |
Tracing the fate of stable isotopically-enriched nutrients is a sophisticated method of describing and quantifying the activity of metabolic pathways. Nuclear Magnetic Resonance (NMR) offers high resolution data, yet is under-utilised due to length of time required to collect the data, quantification requiring multiple samples and complicated analysis. Here we present two techniques, quantitative spectral filters and enhancement of the splitting due to J-coupling in 1H,13C-HSQC NMR spectra, which allow the rapid collection of NMR data in a quantitative manner on a single sample. The reduced duration of HSQC spectra data acquisition opens up the possibility of real-time tracing of metabolism including the study of metabolic pathways in vivo. We show how these novel techniques can be used to trace the fate of labelled nutrients in a whole organ model of kidney preservation prior to transplantation using a porcine kidney as a model organ, and also show how the use of multiple nutrients, differentially labelled with 13C and 15N, can be used to provide additional information with which to profile metabolic pathways. |
url |
https://wellcomeopenresearch.org/articles/3-5/v1 |
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