Determination of acetyl-CoA enrichment in rat heart and skeletal muscle by 1H nuclear magnetic resonance analysis of glutamate in tissue extracts

John G. Jones, Jim Hansen, A. Dean Sherry, Craig R. Malloy, Ronald G. Victor

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

The contribution of a 13C-enriched substrate to the acetyl-CoA pool in animal tissues is typically measured by analysis of glutamate enrichment from tissue extracts. 13C NMR analysis offers the advantages of minimal sample processing and high information content, but has a low analytical sensitivity compared to other methods of tracer analysis such as GC/MS. We present a sensitive, simple, and direct 1H NMR measurement of glutamate C4 enrichment from tissue extracts. The method is demonstrated with heart and hindlimb muscle tissue extracts of rats infused with [2,4,6,8-13C4]octanoate, a source of [2-13C]acetyl-CoA. Glutamate C4 enrichment in extracts of individual hindlimb soleus muscles weighing ≃150 mg and containing approximately 0.3 μmol of glutamate was quantified by 1H NMR within about 40 min. Glutamate C4 enrichment measurements by 1H NMR in heart and gastrocnemius muscle were also highly correlated with independent measurements obtained from 13C NMR isotopomer analysis.

Original languageEnglish (US)
Pages (from-to)201-206
Number of pages6
JournalAnalytical biochemistry
Volume249
Issue number2
DOIs
StatePublished - Jul 1 1997

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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