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Biochemistry (Moscow)

, Volume 74, Issue 4, pp 371–376 | Cite as

Features of mitochondrial energetics in living unicellular eukaryote Tetrahymena pyriformis. A model for study of mammalian intracellular adaptation

  • E. A. Prikhodko
  • I. V. Brailovskaya
  • S. M. Korotkov
  • E. N. MokhovaEmail author
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Abstract

Tetrahymena pyriformis is used in diverse studies as a non-mammalian alternative due to their resemblance in many main metabolic cycles. However, such basic features of mitochondrial energetics as ΔΨ (electrical potential difference across the inner mitochondrial membrane) or maximal stimulation of respiration by uncouplers with different mechanisms of uncoupling, such as DNP (2,4-dinitrophenol) and FCCP (p-trifluoromethoxycarbonylcyanide phenylhydrazone), have not been studied in living ciliates. Tetrahymena pyriformis GL cells during stationary growth phase after incubation under selected conditions were used in this study. Maximal stimulation of cellular respiration by FCCP was about six-fold, thus the proton motive force was high. The DNP uncoupling effect was significantly lower. This suggests low activity of the ATP/ADP-antiporter, which performs not only exchange of intramitochondrial ATP to extramitochondrial ADP, but also helps in the uncoupling process. It participates by a similar mechanism in electrophoretic transport from matrix to cytosol of ATP4− and DNP anion, but not FCCP anion. Thus, in contrast with mammalian mitochondria, T. pyriformis mitochondria cannot rapidly supply the cytosol with ATP; possibly the cells need high intramitochondrial ATP. The difference between DNP and FCCP is hypothetically explained by low ΔΨ value and/or an increase in concentration of long-chain acyl-CoAs, inhibitors of the ATP/ADP-antiporter. The first suggestion is confirmed by absence of mitochondria with bright fluorescence in T. pyriformis stained with the ΔΨ-sensitive probe MitoTracker Red. These data suggest that T. pyriformis cells are useful as a model for study of mitochondrial role in adaptation at the intracellular level.

Key words

ATP/ADP-antiporter membrane potential mitochondria Tetrahymena pyriformis uncouplers DNP intracellular adaptation 

Abbreviations

BSA

bovine serum albumin

DNP

2,4-dinitrophenol

FCCP

p-trifluoromethoxycarbonylcyanide phenylhydrazone

ΔΨ

transmembrane difference of electrical potential across the inner mitochondrial membrane

ΔpH

difference in hydrogen ion concentrations between the two sides of the inner mitochondrial membrane

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • E. A. Prikhodko
    • 1
    • 2
  • I. V. Brailovskaya
    • 3
  • S. M. Korotkov
    • 3
  • E. N. Mokhova
    • 1
    Email author
  1. 1.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia
  2. 2.Faculty of Bioengineering and BioinformaticsLomonosov Moscow State UniversityMoscowRussia
  3. 3.Sechenov Institute of Evolutionary Physiology and BiochemistryRussian Academy of SciencesSt. PetersburgRussia

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