13C isotopomer analysis of glutamate by heteronuclear multiple quantum coherence‐total correlation spectroscopy (HMQC‐TOCSY)

FEBS Letters

Carvalho, Rui A.²  Sherry, A.Dean¹  Malloy, Craig R.³  Jeffrey, F.Mark H.³ 

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

关键词: Indirect detection;    1H Nuclear magnetic resonance;    13C isotopomer analysis;    Two-dimensional nuclear magnetic resonance;    Heteronuclear multiple quantum coherence-total correlation spectroscopy;   

DOI  :  10.1016/S0014-5793(98)01491-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

¹³C has become an important tracer isotope for studies of intermediary metabolism. Information about relative flux through pathways is encoded by the distribution of ¹³C isotopomers in an intermediate pool such as glutamate. This information is commonly decoded either by mass spectrometry or by measuring relative multiplet areas in a ¹³C NMR spectrum. We demonstrate here that groups of glutamate ¹³C isotopomers may be quantified by indirect detection of protons in a 2D HMQC-TOCSY NMR spectrum and that fitting of these data to a metabolic model provides an identical measure of the ¹³C fractional enrichment of acetyl-CoA and relative anaplerotic flux to that given by direct ¹³C NMR analysis. The sensitivity gain provided by HMQC-TOCSY spectroscopy will allow an extension of ¹³C isotopomer analysis to tissue samples not amenable to direct ¹³C detection (∼10 mg soleus muscle) and to tissue metabolites other than glutamate that are typically present at lower concentrations.

【 授权许可】

Unknown