Metabolic profiling by 13C-NMR spectroscopy: [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells

A. Miccheli, A. Tomassini, C. Puccetti, M. Valerio, G. Peluso, F. Tuccillo, M. Calvani, C. Manetti, F. Conti

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Metabolic profiling is defined as the simultaneous assessment of substrate fluxes within and among the different pathways of metabolite synthesis and energy production under various physiological conditions. The use of stable-isotope tracers and the analysis of the distribution of labeled carbons in various intermediates, by both mass spectrometry and NMR spectroscopy, allow the role of several metabolic processes in cell growth and death to be defined. In the present paper we describe the metabolic profiling of Jurkat cells by isotopomer analysis using 13C-NMR spectroscopy and [1,2-13C2]glucose as the stable-isotope tracer. The isotopomer analysis of the lactate, alanine, glutamate, proline, serine, glycine, malate and ribose-5-phosphate moiety of nucleotides has allowed original integrated information regarding the pentose phosphate pathway, TCA cycle, and amino acid metabolism in proliferating human leukemia T cells to be obtained. In particular, the contribution of the glucose-6-phosphate dehydrogenase and transketolase activities to phosphoribosyl-pyrophosphate synthesis was evaluated directly by the determination of isotopomers of the [1′-13C], [4′,5′-13C2]ribosyl moiety of nucleotides. Furthermore, the relative contribution of the glycolysis and pentose cycle to lactate production was estimated via analysis of lactate isotopomers. Interestingly, pyruvate carboxylase and pyruvate dehydrogenase flux ratios measured by glutamate isotopomers and the production of isotopomers of several metabolites showed that the metabolic processes described could not take place simultaneously in the same macrocompartments (cells). Results revealed a heterogeneous metabolism in an asynchronous cell population that may be interpreted on the basis of different metabolic phenotypes of subpopulations in relation to different cell cycle phases.

Original languageEnglish
Pages (from-to)437-448
Number of pages12
JournalBiochimie
Volume88
Issue number5
DOIs
Publication statusPublished - May 2006

Fingerprint

T-Cell Leukemia
T-cells
Metabolism
Nuclear magnetic resonance spectroscopy
Pentoses
Lactic Acid
Magnetic Resonance Spectroscopy
Metabolites
Glucose
Isotopes
Glutamic Acid
Nucleotides
Phosphoribosyl Pyrophosphate
glycyl-prolyl-glutamic acid
Cells
Transketolase
Pyruvate Carboxylase
Fluxes
Mass Spectrometry
Glucosephosphate Dehydrogenase

Keywords

  • C
  • Cancer
  • Metabolic profiling
  • Metabolomics
  • NMR spectroscopy

ASJC Scopus subject areas

  • Biochemistry

Cite this

Metabolic profiling by 13C-NMR spectroscopy : [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells. / Miccheli, A.; Tomassini, A.; Puccetti, C.; Valerio, M.; Peluso, G.; Tuccillo, F.; Calvani, M.; Manetti, C.; Conti, F.

In: Biochimie, Vol. 88, No. 5, 05.2006, p. 437-448.

Research output: Contribution to journalArticle

Miccheli, A, Tomassini, A, Puccetti, C, Valerio, M, Peluso, G, Tuccillo, F, Calvani, M, Manetti, C & Conti, F 2006, 'Metabolic profiling by 13C-NMR spectroscopy: [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells', Biochimie, vol. 88, no. 5, pp. 437-448. https://doi.org/10.1016/j.biochi.2005.10.004
Miccheli, A. ; Tomassini, A. ; Puccetti, C. ; Valerio, M. ; Peluso, G. ; Tuccillo, F. ; Calvani, M. ; Manetti, C. ; Conti, F. / Metabolic profiling by 13C-NMR spectroscopy : [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells. In: Biochimie. 2006 ; Vol. 88, No. 5. pp. 437-448.
@article{94b66cb9fea841f0bd6cf0939101bb9f,
title = "Metabolic profiling by 13C-NMR spectroscopy: [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells",
abstract = "Metabolic profiling is defined as the simultaneous assessment of substrate fluxes within and among the different pathways of metabolite synthesis and energy production under various physiological conditions. The use of stable-isotope tracers and the analysis of the distribution of labeled carbons in various intermediates, by both mass spectrometry and NMR spectroscopy, allow the role of several metabolic processes in cell growth and death to be defined. In the present paper we describe the metabolic profiling of Jurkat cells by isotopomer analysis using 13C-NMR spectroscopy and [1,2-13C2]glucose as the stable-isotope tracer. The isotopomer analysis of the lactate, alanine, glutamate, proline, serine, glycine, malate and ribose-5-phosphate moiety of nucleotides has allowed original integrated information regarding the pentose phosphate pathway, TCA cycle, and amino acid metabolism in proliferating human leukemia T cells to be obtained. In particular, the contribution of the glucose-6-phosphate dehydrogenase and transketolase activities to phosphoribosyl-pyrophosphate synthesis was evaluated directly by the determination of isotopomers of the [1′-13C], [4′,5′-13C2]ribosyl moiety of nucleotides. Furthermore, the relative contribution of the glycolysis and pentose cycle to lactate production was estimated via analysis of lactate isotopomers. Interestingly, pyruvate carboxylase and pyruvate dehydrogenase flux ratios measured by glutamate isotopomers and the production of isotopomers of several metabolites showed that the metabolic processes described could not take place simultaneously in the same macrocompartments (cells). Results revealed a heterogeneous metabolism in an asynchronous cell population that may be interpreted on the basis of different metabolic phenotypes of subpopulations in relation to different cell cycle phases.",
keywords = "C, Cancer, Metabolic profiling, Metabolomics, NMR spectroscopy",
author = "A. Miccheli and A. Tomassini and C. Puccetti and M. Valerio and G. Peluso and F. Tuccillo and M. Calvani and C. Manetti and F. Conti",
year = "2006",
month = "5",
doi = "10.1016/j.biochi.2005.10.004",
language = "English",
volume = "88",
pages = "437--448",
journal = "Biochimie",
issn = "0300-9084",
publisher = "Elsevier",
number = "5",

}

TY - JOUR

T1 - Metabolic profiling by 13C-NMR spectroscopy

T2 - [1,2-13C2]glucose reveals a heterogeneous metabolism in human leukemia T cells

AU - Miccheli, A.

AU - Tomassini, A.

AU - Puccetti, C.

AU - Valerio, M.

AU - Peluso, G.

AU - Tuccillo, F.

AU - Calvani, M.

AU - Manetti, C.

AU - Conti, F.

PY - 2006/5

Y1 - 2006/5

N2 - Metabolic profiling is defined as the simultaneous assessment of substrate fluxes within and among the different pathways of metabolite synthesis and energy production under various physiological conditions. The use of stable-isotope tracers and the analysis of the distribution of labeled carbons in various intermediates, by both mass spectrometry and NMR spectroscopy, allow the role of several metabolic processes in cell growth and death to be defined. In the present paper we describe the metabolic profiling of Jurkat cells by isotopomer analysis using 13C-NMR spectroscopy and [1,2-13C2]glucose as the stable-isotope tracer. The isotopomer analysis of the lactate, alanine, glutamate, proline, serine, glycine, malate and ribose-5-phosphate moiety of nucleotides has allowed original integrated information regarding the pentose phosphate pathway, TCA cycle, and amino acid metabolism in proliferating human leukemia T cells to be obtained. In particular, the contribution of the glucose-6-phosphate dehydrogenase and transketolase activities to phosphoribosyl-pyrophosphate synthesis was evaluated directly by the determination of isotopomers of the [1′-13C], [4′,5′-13C2]ribosyl moiety of nucleotides. Furthermore, the relative contribution of the glycolysis and pentose cycle to lactate production was estimated via analysis of lactate isotopomers. Interestingly, pyruvate carboxylase and pyruvate dehydrogenase flux ratios measured by glutamate isotopomers and the production of isotopomers of several metabolites showed that the metabolic processes described could not take place simultaneously in the same macrocompartments (cells). Results revealed a heterogeneous metabolism in an asynchronous cell population that may be interpreted on the basis of different metabolic phenotypes of subpopulations in relation to different cell cycle phases.

AB - Metabolic profiling is defined as the simultaneous assessment of substrate fluxes within and among the different pathways of metabolite synthesis and energy production under various physiological conditions. The use of stable-isotope tracers and the analysis of the distribution of labeled carbons in various intermediates, by both mass spectrometry and NMR spectroscopy, allow the role of several metabolic processes in cell growth and death to be defined. In the present paper we describe the metabolic profiling of Jurkat cells by isotopomer analysis using 13C-NMR spectroscopy and [1,2-13C2]glucose as the stable-isotope tracer. The isotopomer analysis of the lactate, alanine, glutamate, proline, serine, glycine, malate and ribose-5-phosphate moiety of nucleotides has allowed original integrated information regarding the pentose phosphate pathway, TCA cycle, and amino acid metabolism in proliferating human leukemia T cells to be obtained. In particular, the contribution of the glucose-6-phosphate dehydrogenase and transketolase activities to phosphoribosyl-pyrophosphate synthesis was evaluated directly by the determination of isotopomers of the [1′-13C], [4′,5′-13C2]ribosyl moiety of nucleotides. Furthermore, the relative contribution of the glycolysis and pentose cycle to lactate production was estimated via analysis of lactate isotopomers. Interestingly, pyruvate carboxylase and pyruvate dehydrogenase flux ratios measured by glutamate isotopomers and the production of isotopomers of several metabolites showed that the metabolic processes described could not take place simultaneously in the same macrocompartments (cells). Results revealed a heterogeneous metabolism in an asynchronous cell population that may be interpreted on the basis of different metabolic phenotypes of subpopulations in relation to different cell cycle phases.

KW - C

KW - Cancer

KW - Metabolic profiling

KW - Metabolomics

KW - NMR spectroscopy

UR - http://www.scopus.com/inward/record.url?scp=33745083035&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745083035&partnerID=8YFLogxK

U2 - 10.1016/j.biochi.2005.10.004

DO - 10.1016/j.biochi.2005.10.004

M3 - Article

C2 - 16359766

AN - SCOPUS:33745083035

VL - 88

SP - 437

EP - 448

JO - Biochimie

JF - Biochimie

SN - 0300-9084

IS - 5

ER -