NMR indirect detection of glutamate to measure citric acid cycle flux in the isolated perfused mouse heart

Shawn C. Burgess, Evelyn E. Babcock, F. MarkH Jeffrey, A. Dean Sherry, Craig R. Malloy

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

13C-edited proton nuclear magnetic resonance (NMR) spectroscopy was used to follow enrichment of glutamate C3 and C4 with a temporal resolution of ∼20 s in mouse hearts perfused with 13C-enriched substrates. A fit of the NMR data to a kinetic model of the tricarboxylic acid (TCA) cycle and related exchange reactions yielded TCA cycle (Vtca) and exchange (Vx) fluxes between α-ketoglutarate and glutamate. These fluxes were substrate-dependent and decreased in the order acetate (Vtca = 14.1 μmol g-1 min-1; Vx = 26.5 μmol g-1 min-1) > octanoate (Vtca = 6.0 μmol g-1 min-1; Vx = 16.1 μmol g-1 min-1)lactate (Vtca = 4.2 μmol g-1 min-1; Vx = 6.3 μmol g-1 min-1).

Original languageEnglish (US)
Pages (from-to)163-167
Number of pages5
JournalFEBS Letters
Volume505
Issue number1
DOIs
StatePublished - Sep 7 2001

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Citric Acid Cycle
Glutamic Acid
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Fluxes
Substrates
Nuclear magnetic resonance spectroscopy
Lactic Acid
Acetates
Kinetics
octanoic acid
Proton Magnetic Resonance Spectroscopy

Keywords

  • H nuclear magnetic resonance
  • Cardiac metabolism
  • Indirect detection
  • Tricarboxylic acid cycle kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

NMR indirect detection of glutamate to measure citric acid cycle flux in the isolated perfused mouse heart. / Burgess, Shawn C.; Babcock, Evelyn E.; Jeffrey, F. MarkH; Sherry, A. Dean; Malloy, Craig R.

In: FEBS Letters, Vol. 505, No. 1, 07.09.2001, p. 163-167.

Research output: Contribution to journalArticle

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AU - Jeffrey, F. MarkH

AU - Sherry, A. Dean

AU - Malloy, Craig R.

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N2 - 13C-edited proton nuclear magnetic resonance (NMR) spectroscopy was used to follow enrichment of glutamate C3 and C4 with a temporal resolution of ∼20 s in mouse hearts perfused with 13C-enriched substrates. A fit of the NMR data to a kinetic model of the tricarboxylic acid (TCA) cycle and related exchange reactions yielded TCA cycle (Vtca) and exchange (Vx) fluxes between α-ketoglutarate and glutamate. These fluxes were substrate-dependent and decreased in the order acetate (Vtca = 14.1 μmol g-1 min-1; Vx = 26.5 μmol g-1 min-1) > octanoate (Vtca = 6.0 μmol g-1 min-1; Vx = 16.1 μmol g-1 min-1)lactate (Vtca = 4.2 μmol g-1 min-1; Vx = 6.3 μmol g-1 min-1).

AB - 13C-edited proton nuclear magnetic resonance (NMR) spectroscopy was used to follow enrichment of glutamate C3 and C4 with a temporal resolution of ∼20 s in mouse hearts perfused with 13C-enriched substrates. A fit of the NMR data to a kinetic model of the tricarboxylic acid (TCA) cycle and related exchange reactions yielded TCA cycle (Vtca) and exchange (Vx) fluxes between α-ketoglutarate and glutamate. These fluxes were substrate-dependent and decreased in the order acetate (Vtca = 14.1 μmol g-1 min-1; Vx = 26.5 μmol g-1 min-1) > octanoate (Vtca = 6.0 μmol g-1 min-1; Vx = 16.1 μmol g-1 min-1)lactate (Vtca = 4.2 μmol g-1 min-1; Vx = 6.3 μmol g-1 min-1).

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