Abstract
The isozyme patterns of ovarian malate dehydrogenase (MDH) from various anuran amphibian species were analyzed by isoelectric focusing (IEF). Extensive variability was observed in both the soluble (sMDH) and mitochondrial (mMDH) patterns with as few as two and as many nine bands being visualized in different species. The mean pIs for sMDH ranged from 4.5 to 8.3 and those for mMDH fell between 5.1 and 8.2. The sMDHs are considerably more heat labile in Rana species living in northern latitudes compared to those from southern states. Inhibition with p-chloromercuribenzoate (PCMB) revealed the importance of sulfhydryl groups for the activity of sMDHs, while the functional requirement for these groups in mMDHs appears to be of lesser importance. Observations from these studies lend support to the accumulating evidence that Rana pipiens from such southern locations as New Mexico may have undergone speciation.
Similar content being viewed by others
References
Bailey, G. S., Wilson, A. C., Halver, J., and Johnson, C. (1970). Multiple forms of supernatant malate dehydrogenase in Salmonid fishes: biochemical, immunological and genetic studies. J. Biol. Chem. 2455927.
Baldwin, J., and Aleksiuk, M. (1973). Adaptation of enzymes to temperature: lactate and malate dehydrogenases from platypus and echidna. Comp. Biochem. Physiol. 44B367.
Bisbee, C. A., Baker, M. A., Wilson, A. C., Hadji-Azimi, I., and Fischberg, M. (1977). Albumin phylogeny for clawed frogs (Xenopus). Science 195787.
Brewer, G. J. (1967). Achromatic regions of tetrazolium stained starch gels: inherited electrophoretic variation. Am. J. Human Genet 19(5):674.
Brewer, G. J. (1970). An Introduction to Isozyme Techniques. Academic, New York.
Brown, L. E. (1973). Speciation in Rana pipiens complex. Am. Zool. 1373.
Brown, L. E., and Brown, J. R. (1972). Call types of the Rana pipiens complex. Science 176928.
Chase, J. W., and Dawid, I. B. (1972). Biogenesis of mitochondria during Xenopus laevis development. Develop. Biol. 27504.
Chen, P. W. (1968). Patterns of soluble proteins and multiple forms of dehydrogenases in amphibian development. J. Exp. Zool. 168337.
Davidson, R. G., and Cortner, J. A. (1967). Mitochondrial malate dehydrogenase: a new genetic polymorphism in man. Science 1571569.
Francesconi, R. P., and Villee, C. A. (1968). Changes in malate dehydrogenase isozyme patterns in Arabacia embryos during early development. Comp. Biochem. Physiol. 25747.
Gerding, R. K., and Wolfe, R. G. (1969). Malic dehydrogenase—VIII. Large scale purification and properties of supernatant pig heart enzyme. J. Biol. Chem. 2441164.
Gillespie, H., and Crenshaw, J. W. (1966). Hemoglobin variation in Rana pipiens (Amphibia: Anura). Copeia 1966889.
Grimm, C., and Doherty, D. G. (1961). Properties of the two forms of malic dehydrogenase from beef heart. J. Biol. Chem. 2361980.
Grimwood, B. G., and McDaniel, R. G. (1970). Variant malate dehydrogenase isoenzymes in mitochondrial populations. Biochim. Biophys. Acta 220410.
Hochachka, P. W., and Somero, G. N. (1968). The adaptation of enzymes to temperature. Comp. Biochem. Physiol. 27659.
Hodges, C. T., Wiggins, J. C., and Harrison, J. H. (1977). Investigation of the relation of the pH-dependent dissociation of malate dehydrogenase to modification of the enzyme by N-ethylmaleimide. J. Biol. Chem. 2526038.
Hubby, J. L., and Lewontin, R. C. (1966). A molecular approach to the study of genetic heterozygosity in natural populations—I. The number of alleles at different loci in Drosophila pseudoobscura. Genetics 54577.
Karig, L. M., and Wilson, A. C. (1971). Genetic variation in supernatant malate dehydrogenase of birds and reptiles. Biochem. Genet. 5211.
Kitto, G. B., and Kaplan, N. O. (1966). Purification and properties of chicken heart mitochondrial and supernatant malic dehydrogenases. Biochemistry 123966.
Kitto, G. B., Wasserman, P. M., and Kaplan, N. O. (1966). Enzymatically active conformers of mitochondrial malate dehydrogenase. Proc. Natl. Acad. Sci. U.S.A. 56578.
Kitto, G. B., and Wilson, A. C. (1966). Evolution of malate dehydrogenase in birds. Science 1531408.
Leabeck, D. H. (1975). Isoelectric focusing as an aid to other techniques for the separation and characterization of proteins. In Arbuthnott, J. P., and Beeley, J. P. (eds.), Isoelectric Focusing Butterworths, London, p. 201.
Leese, C. L. (1969). Enzymes and Isoenzymes. In Wolstenholme, G. E. W. and Knight, J. (eds.), Homeostatic Regulators Ciba Found., Churchill Livingstone, Edinburgh.
Lehninger, A. L. (1975). Biochemistry Worth Publishers, New York.
Longo, G. P., and Scandalios, J. G. (1969). Nuclear gene control of mitochondrial malic dehydrogenase in maize. Genetics 62104.
Markert, C. L., and Møller, F. (1959). Multiple forms of enzymes: tissue, ontogenetic, and species specific patterns. Biochemistry 45753.
Massaro, E. J. (1972). Isozyme patterns of Coregonine fishes: evidence for multiple cistrons for lactate and malate dehydrogenases and achromatic bands in the tissues of Prosobium cylindraceum (Pallas) and P. coulteri (Eigenmann and Eigenmann). J. Exp. Zool. 179247.
Massaro, E. J., and Markert, C. L. (1968). Isozyme patterns of Salmonid fishes: evidence for multiple cistrons for lactate dehydrogenase polypeptides. J. Exp. Zool. 168223.
Milkman, R. and Koehler, R. (1976). Isoelectric focusing of MDH and 6-PGDH from Escherichia coli of diverse natural origins. Biochem. Genet. 14517.
Narang, S., and Narang, N. (1974). Characterization of malate dehydrogenase: electrophoresis, isoelectric focusing, thermostability, inhibition and activity studies on homogenates of various organs of Biomphalarice glabrata (Mollusca: Pulmonata). Comp. Biochem. Physiol. 49B477.
O'Brien, S. J. (1973). Comparative analysis of malate dehydrogenases of Drosophila melanogaster. Biochem. Genet. 10191.
Platz, J. E. (1976). Biochemical and morphological variation of leopard frogs in Arizona. Copeia 1976(4):660.
Post, D. D., and Pettus, D. (1966). Variation in Rana pipiens (Anura: Ranidae) of eastern Colorado. Southwest. Nat. 11476.
Righetti, P. G., and Drysdale, J. W. (1976). Isoelectric Focusing North-Holland, Amsterdam.
Seigel, L., and Englard, S. (1961). A comparative study of two malic dehydrogenases from beef heart. Fed. Proc. Fed. Am. Soc. Exp. Biol. 201239.
Shaw, E. R., and Koen, A. L. (1965). On the identity of “nothing dehydrogenase.” J. Histochem. Cytochem. 7431.
Shows, T. B., Chapman, V. M., and Ruddle, F. H. (1970). Mitochondrial malate dehydrogenase and malic enzyme: mendelian inherited electrophoretic variants in the mouse. Biochem. Genet. 4707.
Somero, G. (1969). Enzymic mechanisms of temperature compensation: immediate and evolutionary effects of temperature on enzymes of aquatic poikilotherms. Am. Nat. 103517.
Thorne, C. J. R., Grossman, L. I., and Kaplan, N. O. (1963). Starch gel electrophoresis of malate dehydrogenase. Biochim. Biophys. Acta 73193.
Vesterberg, O. (1975). Comparison and interpretation of complex protein patterns. In P. G. Righetti (ed.), Progress in Isoelectric Focusing and Isotachophoresis North-Holland, Amsterdam, p. 121.
Wall, D. A., and Blackler, A. W. (1974). Enzyme patterns in two species of Xenopus and their hybrids. Develop. Biol. 36379.
Whitt, G. S. (1975). Isoenzymes and developmental biology. In C. L. Markert (ed.), Isozymes, Vol. 3, Academic, New York, p. 1.
Wilson, A. C. (1976). Gene regulation in evolution. In Ayala, F. J. (ed.), Molecular Evolution, Sinauer Associates, Sunderland Press.
Wright, D. A., and Subtelny, S. (1971). Nuclear and cytoplasmic contributions to dehydrogenase phenotypes in hybrid frog embryos. Develop. Biol. 24119.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Webb, A.C., Ingalls, H.M. Analysis of malate dehydrogenase isozymes from anuran amphibian ovary by isoelectric focusing. Biochem Genet 18, 1185–1205 (1980). https://doi.org/10.1007/BF00484347
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00484347