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J. Biol. Chem., Vol. 276, Issue 4, 2586-2599, January 26, 2001
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From the Laboratory of Cardiac Energetics, NHLBI, National
Institutes of Health, Bethesda, Maryland 20892-1061
Parallel activation of heart mitochondria NADH
and ATP production by Ca2+ has been shown to involve
the Ca2+-sensitive dehydrogenases and the
F0F1-ATPase. In the current study we
hypothesize that the response time of Ca2+-activated ATP
production is rapid enough to support step changes in myocardial
workload (~100 ms). To test this hypothesis, the rapid kinetics of
Ca2+ activation of
m
Calcium Activation of Heart Mitochondrial Oxidative
Phosphorylation
RAPID KINETICS OF
m
O2, NADH, AND
LIGHT SCATTERING*
,O2, [NADH], and
light scattering were evaluated in isolated porcine heart mitochondria
at 37 °C using a variety of optical techniques. The addition of
Ca2+ was associated with an initial response time (IRT) of
mO2 that was
dose-dependent with a minimum IRT of 0.27 ± 0.02 s (n = 41) at 535 nM Ca2+. The
IRTs for NADH fluorescence and light scattering in response to
Ca2+ additions were similar to
mO2. The
Ca2+ IRT for
mO2 was
significantly shorter than 1.6 mM ADP (2.36 ± 0.47 s; p 
0.001, n = 13), 2.2 mM Pi (2.32 ± 0.29, p 0.001, n = 13), or 10 mM creatine
(15.6.±1.18 s, p 0.001, n = 18)
under similar experimental conditions. Calcium effects were inhibited with 8 µM ruthenium red (2.4 ± 0.31 s;
p 0.001, n = 16) and reversed with
EGTA (1.6 ± 0.44; p 0.01, n = 6). Estimates of Ca2+ uptake into mitochondria using
optical Ca2+ indicators trapped in the matrix revealed a
sufficiently rapid uptake to cause the metabolic effects observed.
These data are consistent with the notion that extramitochondrial
Ca2+ can modify ATP production, via an increase in matrix
Ca2+ content, rapidly enough to support cardiac work
transitions in vivo.
*
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.
To whom correspondence should be addressed: Laboratory of Cardiac
Energetics, NHLBI, National Institutes of Health, Bldg. 10, Rm.
B1D-400, Bethesda, MD 20892-1061. Tel.: 301-496-2568; Fax:
301-402-2389; E-mail: territop@zeus.nhlbi.nih.gov.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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