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Quantitation of erythrocyte pentose pathway flux with [2-13C]glucose and 1H NMR analysis of the lactate methyl signal

✍ Scribed by Teresa C. Delgado; M. Margarida Castro; Carlos F. Geraldes; John G. Jones

Publisher
John Wiley and Sons
Year
2004
Tongue
English
Weight
84 KB
Volume
51
Category
Article
ISSN
0740-3194

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✦ Synopsis

Abstract

A simple and sensitive NMR method for quantifying excess ^13^C‐enrichment in positions 2 and 3 of lactate by ^1^H NMR spectroscopy of the lactate methyl signal is described. The measurement requires neither signal calibrations nor the addition of a standard and accounts for natural abundance ^13^C‐contributions. As a demonstration, the measurement was applied to ∼3 μmol of lactate generated by erythrocyte preparations incubated with [2‐^13^C]glucose to determine the fraction of glucose metabolized by the pentose phosphate pathway (PP). PP fluxes were estimated from the ratio of excess ^13^C‐enrichment in lactate carbon 3 relative to carbon 2 in accordance with established metabolic models. Under baseline conditions, PP flux accounted for 7 ± 2% of glucose consumption while in the presence of methylene blue, a classical activator of PP activity, its contribution increased to 27 ± 10% of total glucose consumption (P < 0.01). Magn Reson Med 51:1283–1286, 2004. © 2004 Wiley‐Liss, Inc.

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