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2018 ; 3
(ä): 5
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High-Speed Tracer Analysis of Metabolism (HS-TrAM)
#MMPMID29503875
Smith TB
; Patel K
; Munford H
; Peet A
; Tennant DA
; Jeeves M
; Ludwig C
Wellcome Open Res
2018[]; 3
(ä): 5
PMID29503875
show ga
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 (1)H,
(13)C-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
(13)C and (15)N, can be used to provide additional information with which to
profile metabolic pathways.
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