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

doi:10.1006/abio.2000.4930

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 journal › Article › peer-review

26 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 language	English (US)
Pages (from-to)	187-195
Number of pages	9
Journal	Analytical biochemistry
Volume	289
Issue number	2
DOIs	https://doi.org/10.1006/abio.2000.4930
State	Published - Feb 15 2001

Keywords

2D NMR
C isotopomer analysis
H NMR
HSQC
Indirect detection

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1006/abio.2000.4930

Cite this

@article{a69b620f0f4c4689883c10f6e04e70e0,

title = "13C isotopomer analysis of glutamate by J-resolved heteronuclear single quantum coherence spectroscopy",

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.",

keywords = "2D NMR, C isotopomer analysis, H NMR, HSQC, Indirect detection",

author = "Burgess, {Shawn C.} and Carvalho, {Rui A.} and Merritt, {Matthew E.} and Jones, {John G.} and Malloy, {Craig R.} and Sherry, {A. Dean}",

note = "Funding Information: This work was supported by NIH HL-34557 and NIH-RR-02584. We thank Drs. Chris Newgard and Hindrik Mulder for providing the β-cells for the [U-13C]fatty acid experiment.",

year = "2001",

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doi = "10.1006/abio.2000.4930",

language = "English (US)",

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AU - Burgess, Shawn C.

AU - Carvalho, Rui A.

AU - Merritt, Matthew E.

AU - Jones, John G.

AU - Malloy, Craig R.

AU - Sherry, A. Dean

N1 - Funding Information: This work was supported by NIH HL-34557 and NIH-RR-02584. We thank Drs. Chris Newgard and Hindrik Mulder for providing the β-cells for the [U-13C]fatty acid experiment.

PY - 2001/2/15

Y1 - 2001/2/15

N2 - 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.

AB - 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.

KW - 2D NMR

KW - C isotopomer analysis

KW - H NMR

KW - HSQC

KW - Indirect detection

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