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Am J Physiol Endocrinol Metab 266: E1012-E1022, 1994;
0193-1849/94 $5.00
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AJP - Endocrinology and Metabolism, Vol 266, Issue 6 E1012-E1022, Copyright © 1994 by American Physiological Society

ARTICLES

A general model for analysis of the tricarboxylic acid cycle with use of [13C]glutamate isotopomer measurements

J. A. Vogt, A. J. Fischman, M. Kempf, Y. M. Yu, R. G. Tompkins and J. F. Burke
Surgical Services, Massachusetts General Hospital, Boston 02114.

A generalized steady-state model was developed for determining tricarboxylic acid cycle fractional fluxes from 13C nuclear magnetic resonance (NMR) data. The model relates the measured mole fractions of [13C]glutamate isotopomers to the fractional fluxes and predicted mole fractions of isotopomers of oxaloacetate (OAA) and acetyl-CoA. This model includes cycling between OAA and fumarate. Fractional fluxes are determined by fitting the model equations to NMR parameters by use of nonlinear least squares. Although only fractional fluxes can be determined from 13C-NMR data, when they are combined with mass spectroscopic measurements, absolute values can be derived. A specific metabolic system represented by published 13C-NMR data from extracts of hearts perfused with [13C]acetate, [13C]pyruvate (PYR), and [13C]acetate plus [13C]PYR was used to test the model. The intensities of predicted 13C-NMR splitting patterns were compared with observed values, and there was excellent agreement between observed and predicted signal intensities. With this model, important physiological parameters, including the OAA-derived fraction of inflow to PYR, PYR-derived fraction of inflow to acetyl-CoA, citrate-derived fraction of inflow to OAA, and PYR-derived fraction of inflow to OAA, can be determined.


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