13C isotopomer analysis of glutamate by heteronuclear multiple quantum coherence-total correlation spectroscopy (HMQC-TOCSY)

Rui A. Carvalho, F. Mark H Jeffrey, A. Dean Sherry, Craig R. Malloy

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

13C has become an important tracer isotope for studies of intermediary metabolism. Information about relative flux through pathways is encoded by the distribution of 13C isotopomers in an intermediate pool such as glutamate. This information is commonly decoded either by mass spectrometry or by measuring relative multiplet areas in a 13C NMR spectrum. We demonstrate here that groups of glutamate 13C isotopomers may be quantified by indirect detection of protons in a 2D HMQC-TOCSY NMR spectrum and that fitting of these data to a metabolic model provides an identical measure of the 13C fractional enrichment of acetyl-CoA and relative anaplerotic flux to that given by direct 13C NMR analysis. The sensitivity gain provided by HMQC-TOCSY spectroscopy will allow an extension of 13C isotopomer analysis to tissue samples not amenable to direct 13C detection (~10 mg soleus muscle) and to tissue metabolites other than glutamate that are typically present at lower concentrations. Copyright (C) 1998 Federation of European Biochemical Societies.

Original languageEnglish (US)
Pages (from-to)382-386
Number of pages5
JournalFEBS Letters
Volume440
Issue number3
DOIs
StatePublished - Dec 4 1998

Fingerprint

Glutamic Acid
Spectrum Analysis
Spectroscopy
Nuclear magnetic resonance
Tissue
Fluxes
Acetyl Coenzyme A
Metabolites
Metabolism
Isotopes
Nuclear magnetic resonance spectroscopy
Mass spectrometry
Muscle
Protons
Mass Spectrometry
Skeletal Muscle
Magnetic Resonance Spectroscopy
Muscles
Carbon-13 Magnetic Resonance Spectroscopy

Keywords

  • C isotopomer analysis
  • H Nuclear magnetic resonance
  • Heteronuclear multiple quantum coherence-total correlation spectroscopy
  • Indirect detection
  • Two-dimensional nuclear magnetic resonance

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

13C isotopomer analysis of glutamate by heteronuclear multiple quantum coherence-total correlation spectroscopy (HMQC-TOCSY). / Carvalho, Rui A.; Jeffrey, F. Mark H; Sherry, A. Dean; Malloy, Craig R.

In: FEBS Letters, Vol. 440, No. 3, 04.12.1998, p. 382-386.

Research output: Contribution to journalArticle

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AU - Malloy, Craig R.

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N2 - 13C has become an important tracer isotope for studies of intermediary metabolism. Information about relative flux through pathways is encoded by the distribution of 13C isotopomers in an intermediate pool such as glutamate. This information is commonly decoded either by mass spectrometry or by measuring relative multiplet areas in a 13C NMR spectrum. We demonstrate here that groups of glutamate 13C isotopomers may be quantified by indirect detection of protons in a 2D HMQC-TOCSY NMR spectrum and that fitting of these data to a metabolic model provides an identical measure of the 13C fractional enrichment of acetyl-CoA and relative anaplerotic flux to that given by direct 13C NMR analysis. The sensitivity gain provided by HMQC-TOCSY spectroscopy will allow an extension of 13C isotopomer analysis to tissue samples not amenable to direct 13C detection (~10 mg soleus muscle) and to tissue metabolites other than glutamate that are typically present at lower concentrations. Copyright (C) 1998 Federation of European Biochemical Societies.

AB - 13C has become an important tracer isotope for studies of intermediary metabolism. Information about relative flux through pathways is encoded by the distribution of 13C isotopomers in an intermediate pool such as glutamate. This information is commonly decoded either by mass spectrometry or by measuring relative multiplet areas in a 13C NMR spectrum. We demonstrate here that groups of glutamate 13C isotopomers may be quantified by indirect detection of protons in a 2D HMQC-TOCSY NMR spectrum and that fitting of these data to a metabolic model provides an identical measure of the 13C fractional enrichment of acetyl-CoA and relative anaplerotic flux to that given by direct 13C NMR analysis. The sensitivity gain provided by HMQC-TOCSY spectroscopy will allow an extension of 13C isotopomer analysis to tissue samples not amenable to direct 13C detection (~10 mg soleus muscle) and to tissue metabolites other than glutamate that are typically present at lower concentrations. Copyright (C) 1998 Federation of European Biochemical Societies.

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