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

FEBS Letters

Sherry, A.Dean¹  Babcock, Evelyn E.²  Burgess, Shawn C.¹  Malloy, Craig R.²  Jeffrey, F.Mark H.² 

[1] Department of Chemistry, University of Texas at Dallas, Dallas, TX, USA;The Mary Nell and Ralph B. Rogers Magnetic Resonance Center, Department of Radiology, University of Texas Southwestern Medical Center at Dallas, 5801 Forest Park Road, Dallas, TX 75235-9085, USA

关键词: Indirect detection;    1H nuclear magnetic resonance;    Tricarboxylic acid cycle kinetics;    Cardiac metabolism;   

DOI  :  10.1016/S0014-5793(01)02799-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 (V _tca) and exchange (V _x) fluxes between α-ketoglutarate and glutamate. These fluxes were substrate-dependent and decreased in the order acetate (V _tca=14.1 μmol g⁻¹ min⁻¹; V _x=26.5 μmol g⁻¹ min⁻¹)>octanoate (V _tca=6.0 μmol g⁻¹ min⁻¹; V _x=16.1 μmol g⁻¹ min⁻¹)>lactate (V _tca=4.2 μmol g⁻¹ min⁻¹; V _x=6.3 μmol g⁻¹ min⁻¹).

【 授权许可】

Unknown