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Histochemical modification of the active site of succinate dehydrogenase withN-acetylimidazole

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Summary

The kinetics of acetylation of mitochondrial succinate dehydrogenase [EC 1.3.99.1] in the two fibre types (A and C) of rat gastrocnemius withN-acetylimidazole was studied by a newly modified histochemical technique.

  1. 1.

    Acetylimidazole partially inactivated the enzyme, but subsequent deacetylation with hydroxylamine restored the enzyme activity completely.

  2. 2.

    Inactivation of the enzyme by acetylimidazole was prevented by malonate, which is a competitive inhibitor of the enzyme.

  3. 3.

    The value of the inhibition constant (Ki=34µM) for malonate, obtained from the dependence of the pseudo-first order rate constant of acetylation of the enzyme with acetylimidazole on the malonate concentration, was in good agreement with theKi value (33µM) obtained by a different method, the dependence of the initial velocity of succinate oxidation by the dehydrogenase on the substrate concentration in the presence of malonate.

These findings suggest that a tyrosyl residue is located in the malonate binding site (the active site) of succinate dehydrogenase in the gastrocnemius and plays a role in substrate binding, but is not a catalytic group.

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Authors and Affiliations

  1. Department of Oral Anatomy, School of Dentistry, Tokushima University, Kuramoto-cho, 770, Tokushima, Japan

    Yoshiko Nakae

  2. General Laboratory for Medical Research, School of Medicine, Tokushima University, Kuramoto-cho, 770, Tokushima, Japan

    Masayuki Shono

Authors
  1. Yoshiko Nakae
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  2. Masayuki Shono
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Nakae, Y., Shono, M. Histochemical modification of the active site of succinate dehydrogenase withN-acetylimidazole. Histochem J 18, 169–174 (1986). https://doi.org/10.1007/BF01676117

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  • DOI: https://doi.org/10.1007/BF01676117

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