Molecular and Cellular Biochemistry

, Volume 174, Issue 1–2, pp 43–51 | Cite as

Experimental evidence for dynamic compartmentation of ADP at the mitochondrial periphery: Coupling of mitochondrial adenylate kinase and mitochondrial hexokinase with oxidative phosphorylation under conditions mimicking the intracellular colloid osmotic pressure

  • Fanny Dorine Laterveer
  • Klaas Nicolay
  • Frank Norbert Gellerich
Article

Abstract

Dextran M20 was added to isolated rat liver mitochondria to mimic cytosolic macromolecules. Under these conditions, the morphological changes in the mitochondrial periphery that occur upon isolation of the organelle are restored, i.e. the volume of the intermembrane space decreases and the contact site frequency increases. The ADP routing from mitochondrial kinases at various locations was investigated by using the activities of oxidative phosphorylation and externally added pyruvate kinase as sensors for ADP transport into the matrix and extramitochondrial compartment, respectively. The studies reveal that a significant fraction of the ADP generated by either adenylate kinase in the intermembrane space or by outer membrane bound hexokinase isozyme I, is not accessible to extramitochondrial pyruvate kinase. Quantitative information on the ADP compartmentation in rat liver mitochondria was obtained by comparing the ADP supply from mitochondrial kinases to oxidative phosphorylation with that of non-bound, extramitochondrially located kinases. This approach allowed us to estimate the ADP diffusion gradients which were present across the outer membrane and between the compartment formed by bound hexokinase and the extramitochondrial compartment. In the presence of 10% dextran M20 these ADP gradients amounted to approximately 12 µM. The possible role of mitochondrial kinases in ADP transport into mitochondria in vivo is discussed. (Mol Cell Biochem 174: 43–51, 1997)

macromolecules oxidative phosphorylation adenylate kinase hexokinase compartmentation concentration gradients 

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Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Fanny Dorine Laterveer
    • 1
    • 2
  • Klaas Nicolay
    • 1
  • Frank Norbert Gellerich
    • 1
  1. 1.Department of in vivo NMR Spectroscopy, Bijvoet Center for Biomolecular ResearchUtrecht UniversityThe Netherlands
  2. 2.Center for Biomembranes and Lipid Enzymology, Department of Biochemistry of MembranesUtrecht UniversityThe Netherlands

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