Cardiac creatine kinase metabolite compartments revealed by NMR magnetization transfer spectroscopy and subcellular fractionation

Biochemistry
Frederic JoubertJacqueline A Hoerter

Abstract

In the perfused rat heart NMR inversion transfer revealed the existence of a compartment of ATP not exchanging through creatine kinase (CK), as demonstrated by an apparent discrepancy between the forward (F(f)) and reverse (F(r)) CK flux if this compartment was neglected in the analysis [Joubert et al. (2000) Biophys. J. 79, 1-13]. To localize this compartment, CK fluxes were measured by inversion of PCr (inv-PCr) or gamma ATP (inv-ATP), and the distribution of metabolites between mitochondria and cytosol was studied by subcellular fractionation. Physiological conditions were designed to modify the concentration and distribution of CK metabolites (control, adenylate depletion, inhibition of respiration, KCl arrest). Depending on cardiac activity, mitochondrial ATP (mito-ATP) assessed by fractionation varied from 11% to 30% of total ATP. In addition, the apparent flux discrepancy increased together with mito-ATP (F(f)/F(r) ranged from 0.85 to 0.50 in inv-PCr and from 1.13 to 1.88 in inv-ATP). Under conditions masking the influence of the ATP-P(i) exchange on CK flux, the ATP compartment could be directly quantified by the apparent flux discrepancy; its size was similar to that of mito-ATP measured by fractionation. Thus NMR inve...Continue Reading

References

Jul 1, 1975·Journal of Molecular and Cellular Cardiology·R A AltschuldG P Brierley
Aug 1, 1988·The American Journal of Physiology·J A HoerterM Guéron
May 1, 1984·Journal of Molecular and Cellular Cardiology·G K Asimakis, V R Conti
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Apr 7, 1980·FEBS Letters·R A KauppinenI E Hassinen
Nov 14, 1997·The American Journal of Physiology·V StepanovJ A Hoerter

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Citations

Sep 28, 2010·Biochimica Et Biophysica Acta·Renée Ventura-ClapierFrédéric Joubert
Jul 28, 2016·Magnetic Resonance in Medicine : Official Journal of the Society of Magnetic Resonance in Medicine·Bertrand PouymayouChris Boesch
Mar 12, 2002·The Journal of Biological Chemistry·Frederic JoubertJacqueline A Hoerter
Nov 3, 2020·Frontiers in Cell and Developmental Biology·Jérôme PiquereauRenée Ventura-Clapier

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