Electrophoretic thermostability tests of soluble malate dehydrogenases (sMDH) isozymes in tissue extracts of 21 subtropical fish belonging to the orders Characiformes, Siluriformes and Perciformes showed three distinct results. The first, characterized by thermal stability of the slowest-migrating band or A-isoform, was detected in 52% of all species. The second, exhibited in 29% of the species analyzed, had a bidirectionally divergent pattern of their sMDH locus expression, and was characterized by a nondivergent thermostability pattern of both sMDH-A* and B*. In the third category, obtained in 19% of the species studied (the four Siluriformes species), thermostability of the fastest-migrating bands, or B-isoforms, was observed. Comparison of the effects of habitat temperature on the activity of paralogous and orthologous isoforms in tissue extracts of two of these species with different thermostability properties (Leporinus friderici - thermostable sMDH-A*, and Pimelodus maculatus - reverse thermostability properties or reverse electrophoretic pattern), collected during winter and summer months, showed that A and B subunits were present at different quantitative levels and their activities were nearly season independent. Differences in susceptibility to temperature (50°C) of both sMDH loci from tissue extracts of these species were found. In P. maculatus, these susceptibilities helped strengthen one of the hypotheses: the reverse thermostability pattern, where the fastest-migrating band or the B-isoform was the thermostable sMDH. Thus, temperature differences among orthologous homologues of sMDH seem to have occurred in these acclimatized species, where the fastest-migrating band, usually muscle specific and thermolabile in most teleosts, appeared in P. maculatus as the thermostable isoform.
No presente trabalho, mostramos em ortólogos da sMDH de 21 espécies de peixes subtropicais das ordens Characiformes, Siluriformes e Perciformes, três diferentes estabilidades térmicas. A primeira, caracterizada pela termoestabilidade do componente menos anódico ou isoforma-A, foi detectada em 52% de todas as espécies. A segunda, exibida por 29% das espécies aqui analisadas, caracterizou-se por um padrão não-divergente de termoestabilidade dos locos sMDH-A* e sMDH-B*. Na terceira resposta, obtida em 19% das espécies analisadas (as 4 espécies Siluriformes), foi observada a termoestabilidade da banda mais anódica ou isoforma-B. O efeito da temperatura ambiental na atividade relativa de isoformas parálogas e ortólogas de duas dessas espécies com diferentes respostas térmicas (Leporinus friderici - sMDH-A*, termoestável, e Pimelodus maculatus, termoestabilidade reversa ou padrão eletroforético reverso), coletadas em meses de inverno e de verão, mostrou que as subunidades A e B estão presentes em seus extratos de tecidos em diferentes níveis quantitativos e suas atividades relativas são, praticamente, independentes da época de coleta. Na incubação de extratos de tecidos dessas 2 espécies a 50°C, diferentes respostas de inativa...

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A recent locus duplication hypothesis for sMDH-B* was proposed to explain the complex electrophoretic pattern of six bands detected for the soluble form of malate dehydrogenase (MDH, EC 1.1.1.37) in 84% of the Geophagus brasiliensis (Cichlidae, Perciformes) analyzed (AB1B2 individuals). Klebe's serial dilutions were carried out in skeletal muscle extracts. B1 and B2 subunits had the same visual end-points, reflecting a nondivergent pattern for these B-duplicated genes. Since there is no evidence of polyploidy in the Cichlidae family, MDH-B* loci must have evolved from regional gene duplication. Tissue specificities, thermostability and kinetic tests resulted in similar responses from both B-isoforms, in both sMDH phenotypes, suggesting that these more recently duplicated loci underwent the same regulatory gene action. Similar results obtained with the two sMDH phenotypes did not show any indication of a six-banded specimen adaptive advantage in subtropical regions.
A fim de explicar o padrão eletroforético de seis componentes detectado para a malato desidrogenase solúvel (MDH, EC 1.1.1.37) em 84% dos exemplares de G. brasiliensis analisados (Cichlidae, Perciformes), uma duplicação recente no loco sMDH-B* é sugerida. Diluições seriadas de Klebe realizadas com extratos de músculo esquelético mostraram para as subunidades B1 e B2 o mesmo ponto final visual sugerindo um padrão de expressão não divergente para esses genes duplicados. Uma vez que não existe evidência de poliploidia na família Cichlidae, é sugerido que a duplicação no loco sMDH-B* seja resultante de uma duplicação regional. Especificidade tissular, termoestabilidade e propriedades cinéticas mostraram-se similares para as isoformas B, em ambos os fenótipos detectados, sugerindo estarem esses sob a ação do mesmo gene regulador. Os resultados similares obtidos para os fenótipos de três (AB1) e seis (AB1B2) componentes aqui analisados não mostraram nenhum indicativo de vantagem adaptativa deste último sobre o primeiro, em região subtropical

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Kinetic properties and thermal stabilities of Hoplias malabaricus liver and skeletal muscle unfractionated malate dehydrogenase (MDH, EC 1.1.1.37) and its isolated isoforms were analyzed to further study the possible sMDH-A* locus duplication evolved from a recent tandem duplication. Both A (A1 and A2) and B isoforms had similar optima pH (7.5-8.0). While Hoplias A isoform could not be characterized as thermostable, B could as thermolabile. A isoforms differed from B isoform in having higher Km values for oxaloacetate. The possibly duplicated A2 isoform showed higher substrate affinity than the A1. Hoplias duplicated A isoforms may influence the direction of carbon flow between glycolisis and gluconeogenesis.