13C isotopomer analysis of glutamate by J-resolved heteronuclear single quantum coherence spectroscopy

Shawn C. Burgess, Rui A. Carvalho, Matthew E. Merritt, John G. Jones, Craig R. Malloy, A. Dean Sherry

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

13C NMR isotopomer analysis is a powerful method for measuring metabolic fluxes through pathways intersecting in the tricarboxylic acid cycle. However, the inherent insensitivity of 13C NMR spectroscopy makes application of isotopomer analysis to small tissue samples (mouse tissue, human biopsies, or cells grown in tissue culture) problematic. 1H NMR is intrinsically more sensitive than 13C NMR and can potentially supply the same information via indirect detection of 13C providing that isotopomer information can be preserved. We report here the use of J-resolved HSQC (J-HSQC) for 13C isotopomer analysis of tissue samples. We show that J-HSQC reports isotopomer multiplet patterns identical to those reported by direct 13C detection but with improved sensitivity.

Original languageEnglish (US)
Pages (from-to)187-195
Number of pages9
JournalAnalytical Biochemistry
Volume289
Issue number2
DOIs
Publication statusPublished - Feb 15 2001

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Keywords

  • 2D NMR
  • C isotopomer analysis
  • H NMR
  • HSQC
  • Indirect detection

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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