Mathematical modelling for the generation of L-[3-2H,3-13C]lactic acid isotopomers by erythrocytes exposed to either D-[1-13C]glucose or D-[6-13C]glucose in the presence of2H2O

Malaisse, Willy; Biesemans, Monique

doi:10.1007/bf01457394

Cited by 7 publications

(1 citation statement)

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“…In order to further characterize the metabolism of o-glucose in RINm5F cells, the experimental data were analyzed according to a model described elsewhere (15). This model takes into account the relative amounts of each C,-deuterated isotopomer after a single or dual backand-forth interconversion of pyruvate and L-alanine (Table 4).…”

Section: Modelingmentioning

confidence: 99%

¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

Biesemans¹,

Kayser²,

Malaisse

1994

Magnetic Resonance in Med

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Tumoral pancreatic islet cells of the RIN5mF line were incubated for 120 min in media prepared in 2H2O and containing D-[1-13C]glucose, D-[2-13C]glucose, and D-[6-13C]glucose. The generation of C2- and C3-deuterated lactic acid was assessed by 13C NMR. The interpretation of experimental results suggests that a) the efficiency of deuteration on the C1 of D-fructose 6-phosphate does not exceed about 47% and 4% in the phosphoglucoisomerase and phosphomannoisomerase reactions, respectively; b) approximately 38% of the molecules of D-glyceraldehyde 3-phosphate generated from D-glucose escape deuteration in the sequence of reactions catalyzed by triose phosphate isomerase and aldolase; and c) about 41% of the molecules of pyruvate generated by glycolysis are immediately converted to lactate, the remaining 59% of pyruvate molecules undergoing first a single or double back-and-forth interconversion with L-alanine. It is proposed that this methodological approach, based on high resolution 13C NMR spectroscopy, may provide novel information on the regulation of back-and-forth interconversion of glycolytic intermediates in intact cells as modulated, for instance, by enzyme-to-enzyme tunneling.

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Section: Modelingmentioning

confidence: 99%

¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

Biesemans¹,

Kayser²,

Malaisse

1994

Magnetic Resonance in Med

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Isotopic methods for probing organization of cellular metabolism

Sherry¹,

Malloy²

1996

Cell Biochemistry & Function

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These examples serve to illustrate that it is now possible to investigate metabolism in intact tissues using a variety of biophysical methods. While we have concentrated on NMR methods, reflecting our own interests in using 13C as a metabolic tracer, GC-mass spectroscopy can often provide similar metabolic information and has the advantage of increased sensitivity over NMR. Combining either or both of these technologies with cleaver 'chemical biopsy' methods offers new opportunities to examine what may seem to be old metabolic questions in a much more relevant environment, the native state.

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A computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints: Analysis of overlapping ¹³C‐²H multiplets of ¹³C enriched metabolites from cell suspensions incubated in deuterated media

Laatikainen¹,

Niemitz²,

Malaisse³

et al. 1996

Magnetic Resonance in Med

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108

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A computational strategy for the deconvolution of complex spectra involving scalar multiplet patterns is presented. This approach fits spectra that can be composed of single resonances as well as scalar coupling multiplets for which resonance frequencies, intensities, and lineshape parameters can be optimized. For multiplets, the coupling constant also is optimized. Any external information about the optimizable parameters can be taken into account as external constraints. A lineshape described by absorptive and dispersive Lorentzian and Gaussian contributions and the baseline with up to 40 Fourier and polynomial terms can likewise be optimized. The effectiveness of the procedure is assessed on the basis of computer simulated deconvolutions of a composite of 1J(13C-2H) multiplets arising from a mixture of all possible 13C-2H isotopomers of deuterated L-[3-13C]lactate generated from cell preparations incubated with D-[1-13C]glucose in D2O, which was analyzed previously with a manual deconvolution procedure (R. Willem, M. Biesemans, F. Kayser, W. J. Malaisse, Magn, Reson. Med. 31, 259-267 (1994)). The use of constraints is shown to lead to an improvement in the results. The fitting strategies and the importance of the baseline as an origin of bias are discussed.

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Mathematical modelling for the generation of L-[3-2H,3-13C]lactic acid isotopomers by erythrocytes exposed to either D-[1-13C]glucose or D-[6-13C]glucose in the presence of2H2O

Cited by 7 publications

References 7 publications

¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

Isotopic methods for probing organization of cellular metabolism

A computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints: Analysis of overlapping ¹³C‐²H multiplets of ¹³C enriched metabolites from cell suspensions incubated in deuterated media

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Mathematical modelling for the generation of L-[3-2H,3-13C]lactic acid isotopomers by erythrocytes exposed to either D-[1-13C]glucose or D-[6-13C]glucose in the presence of2H2O

Cited by 7 publications

References 7 publications

13C NMR study of the generation of C2‐ and C3,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐13C]‐, D‐[2‐13C]‐ and D‐[6‐13C]‐Glucose in 2H2O

13C NMR study of the generation of C2‐ and C3,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐13C]‐, D‐[2‐13C]‐ and D‐[6‐13C]‐Glucose in 2H2O

Isotopic methods for probing organization of cellular metabolism

A computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints: Analysis of overlapping 13C‐2H multiplets of 13C enriched metabolites from cell suspensions incubated in deuterated media

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¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

¹³C NMR study of the generation of C₂‐ and C₃,‐deuterated lactic acid by tumoral pancreatic islet cells exposed to D‐[1‐¹³C]‐, D‐[2‐¹³C]‐ and D‐[6‐¹³C]‐Glucose in ²H₂O

A computational strategy for the deconvolution of NMR spectra with multiplet structures and constraints: Analysis of overlapping ¹³C‐²H multiplets of ¹³C enriched metabolites from cell suspensions incubated in deuterated media