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J. Biol. Chem., Vol. 277, Issue 21, 18469-18476, May 24, 2002
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From INSERM U-446, Cardiologie Cellulaire et Moléculaire,
Université Paris-Sud, Faculté de Pharmacie, 92296 Chatenay Malabry, France
The subcellular fluxes of exchange of ATP and
phosphocreatine (PCr) between mitochondria, cytosol, and ATPases were
assessed by 31P NMR spectroscopy to investigate the
pathways of energy transfer in a steady state beating heart. Using a
combined analysis of four protocols of inversion magnetization transfer
associated with biochemical data, three different creatine kinase (CK)
activities were resolved in the rat heart perfused in isovolumic
control conditions: (i) a cytosolic CK functioning at equilibrium
(forward, Ff = PCr This work is dedicated to W. E. Jacobus, who initiated the
interest (of J. A. H.) in CK and NMR.
31P NMR Detection of Subcellular Creatine Kinase
Fluxes in the Perfused Rat Heart
CONTRACTILITY MODIFIES ENERGY TRANSFER PATHWAYS*
,
ATP, and reverse flux,
Fr = ATP PCr = 3.3 mM·s
1), (ii) a CK localized in the vicinity
of ATPases (MM-CK bound isoform) favoring ATP synthesis
(Ff = 1.7 × Fr), and
(iii) a mitochondrial CK displaced toward PCr synthesis
(Ff = 0.3 and Fr = 2.6 mM·s1). This study thus provides the first
experimental evidence that the energy is carried from mitochondria to
ATPases by PCr (i.e. CK shuttle) in the whole heart. In
contrast, a single CK functioning at equilibrium was sufficient to
describe the data when ATP synthesis was partly inhibited by cyanide
(0.15 mM). In this case, ATP was directly transferred from
mitochondria to cytosol suggesting that cardiac activity modified
energy transfer pathways. Bioenergetic implications of the localization
and activity of enzymes within myocardial cells are discussed.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a grant from the French Ministère de la Recherche.
§
To whom correspondence should be addressed: U-446 INSERM,
Cardiologie Cellulaire et Moléculaire, Faculté de
Pharmacie, 5 rue J.B. Clément, 92296 Chatenay-Malabry, France.
Tel.: 33-1-46835759; Fax: 33-1-46835475; E-mail:
jacqueline.hoerter@cep.u-psud.fr.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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