The multiple soluble malate dehydrogenase of Hoplias malabaricus (Erythrinidae, Characiformes)

  • M. R. Aquino-Silva
  • M. L. B. Schwantes
  • A. R. Schwantes
Part of the EBO — Experimental Biology Online Annual book series (EBOEXP, volume 1996/1997)


To explain the six-banded pattern obtained upon electrophoresis of the soluble form of malate dehydrogenase (sMDH, EC from the characiform Hoplias malabaricus, a recent locus duplication of its A isoform (sMDH-A*), in addition to its sMDH-B* isoform, was proposed. Klebe’s serial dilutions carried out using skeletal muscle, heart and liver extracts showed that the A1 and A2 subunits have the same visual end-points, indicating that these A-duplicated genes have a nondivergent pattern. Since there is no evidence of polyploidy in the Erythrinidae family, the MDH-A* loci have probably evolved from regional gene duplication. While these sMDH-A* loci encode nondivergent thermostable isoforms, the sMDH-B* encodes a thermolabile one. Thermostable sMDHs differ from the thermolabile sMDHs in that they have a higher K m of oxaloacetate. Liver, muscle and heart unfractionated sMDH levels at three different temperature and two pH regimens were analysed and the results showed that, in the adaptative temperature range of Hoplias, the variation in K m under conditions of constant pH (imidazole buffer) was less (approximately threefold) than that measured in the presence of temperature-dependent pH imidazole buffer (sevenfold). Estimation of the ratio of both isoforms in these tissues by Klebe’s method showed that, in unfractionated liver — where K m values were the highest and the minimum K m was obtained at 30°C (both for temperature-dependent pH and constant-pH imidazole buffer) — the duplicate A (thermostable, A1 and A2) and B (thermolabile) subunits were detected in a ratio of 2:1. On the other hand, in muscle extracts — in which the lowest K m values were measured, with the minimum K m at 10–20°C (temperature-dependent pH and constant-pH imidazole buffer, respectively) — a ratio of two thermolabile to one thermostable subunits was observed.

Key words

Characiformes Hoplias malabaricus Km Recent locus duplication Soluble malate dehydrogenase Thermolabile isoform Thermostable isoform 


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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • M. R. Aquino-Silva
    • 1
  • M. L. B. Schwantes
    • 1
  • A. R. Schwantes
    • 1
  1. 1.Department of Genetics and Evolution, Laboratory for Molecular EvolutionUniversidade Federal de São CarlosS. Carlos, SPBrazil

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