Modular kinetic analysis of the adenine nucleotide translocator-mediated effects of palmitoyl-CoA on the oxidative phosphorylation in isolated rat liver mitochondria.
To test whether long-chain fatty acyl-CoA esters link obesity with type 2 diabetes through inhibition of the mitochondrial adenine nucleotide translocator, we applied a system-biology approach, dual modular kinetic analysis, with mitochondrial membrane potential (Deltapsi) and the fraction of matrix ATP as intermediates. We found that 5 mumol/l palmitoyl-CoA inhibited adenine nucleotide translocator, without direct effect on other components of oxidative phosphorylation. Indirect effects depended on how oxidative phosphorylation was regulated. When the electron donor and phosphate acceptor were in excess, and the mitochondrial "work" flux was allowed to vary, palmitoyl-CoA decreased phosphorylation flux by 38% and the fraction of ATP in the medium by 39%. Deltapsi increased by 15 mV, and the fraction of matrix ATP increased by 46%. Palmitoyl-CoA had a stronger effect when the flux through the mitochondrial electron transfer chain was maintained constant: Deltapsi increased by 27 mV, and the fraction of matrix ATP increased 2.6 times. When oxidative phosphorylation flux was kept constant by adjusting the rate using hexokinase, Deltapsi and the fraction of ATP were not affected. Palmitoyl-CoA increased the extramitochondrial AMP concentration significantly. The effects of palmitoyl-CoA in our model system support the proposed mechanism linking obesity and type 2 diabetes through an effect on adenine nucleotide translocator.
Ciapaite
Jolita
J
Department of Molecular Cell Physiology, Institute for Molecular Cell Biology, BioCenter Amsterdam, Faculty of EarthLife Sciences, Vrije Universiteit, De Boelelaan 1085, NL-1081 HV Amsterdam, the Netherlands.
Van Eikenhorst
Gerco
G
Bakker
Stephan J L
SJ
Diamant
Michaela
M
Heine
Robert J
RJ
Wagner
Marijke J
MJ
Westerhoff
Hans V
HV
Krab
Klaas
K
D000818:Animals; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008930:Mitochondria, Liver / Q000378:metabolism*; D000226:Mitochondrial ADP, ATP Translocases / Q000502:physiology*; D010085:Oxidative Phosphorylation; D010171:Palmitoyl Coenzyme A / Q000502:physiology*; D051381:Rats
D010171:Palmitoyl Coenzyme A; D000226:Mitochondrial ADP, ATP Translocases