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Helgoländer Meeresuntersuchungen

, Volume 46, Issue 4, pp 435–445 | Cite as

Mitochondrial ATPase in the gills of the shore crabCarcinus maenas

  • D. Siebers
  • J. Hentschel
  • K. Böttcher
  • C. Lucu
Article

Abstract

Posterior gills (No. 7 and 8) of shore crabsCarcinus maenas were homogenized and fractionated by means of differential and density gradient centrifugation. Employment of marker enzymes Na-K-ATPase and carbonic anhydrase for plasma membranes and cytochrome oxidase for mitochondria showed that these structural elements were separated. Ultramicroscopic investigations of combined fractions confirmed the presence of the respective mitochondrial and vesicular plasma membrane structures. An ATPase which did not depend on the presence of sodium (20 mM) ions in the incubation medium but on the presence of potassium (20 mM) ions only was found in the mitochondrial fractions. The mitochondrial ATPase was tightly bound to cellular particulates and activated approximately threefold by bicarbonate (20 mM) ions. The activity of this ATPase was nearly completely inhibited by oligomycin (1 μg ml−1) and greatly inhibited by low levels (5 mM) of thiocyanate and calcium ions, the Ki for Ca2+ being ca 4 mM. The results obtained confirm literature data on high mitochondrial densities in crab gills and allow the assumption of significant rates of energy metabolism in these organs. Considering its properties the mitochondrial ATPase is clearly distinct from crab gill Na-K-ATPase and can be measured specifically in samples containing Na-K-ATPase. Mitochondrial ATPase is therefore considered a suitable and reliable marker enzyme for mitochondria.

Keywords

Carbonic Anhydrase Thiocyanate Incubation Medium Cytochrome Oxidase Gradient Centrifugation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Biologische Anstalt Helgoland 1992

Authors and Affiliations

  • D. Siebers
    • 1
  • J. Hentschel
    • 2
  • K. Böttcher
    • 1
  • C. Lucu
    • 3
  1. 1.Biologische Anstalt HelgolandHamburg 52Federal Republic of Germany
  2. 2.Fakultät für BiologieUniversität KonstanzKonstanz 1Federal Republic of Germany
  3. 3.Center for Marine ResearchRuder Boskovic InstituteBovinjCroatia

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