Summary
When apparent Michaelis constants (K m's) for pyruvate of M4-lactate dehydrogenases from differently thermally adapted vertebrates are measured at the species' normal cell temperatures, a marked degree of conservation inK m is observed, but only when the pH of the assay medium is varied in the manner in which intracellular pH varies with temperature in most animals (Fig. 2).K m measurements performed at a constant pH do not yield this high degree of interspecific conservation inK m (Figs. 2 and 3).
The temperature dependence of intracellular pH preserves the charge states of imidazoles of protein histidines during temperature transitions. Thus under intracellular conditions the ionization state of the active site histidine of LDH will be independent of temperature, reducing the temperature dependence of pyruvate binding. This effect appears important in the contexts of short-term temperature variation experienced by an individual ectotherm and of long-term, evolutionary temperature changes important in speciation processes.
These findings emphasize the importance of utilizing biologically realistic pH values in enzyme studies if major adaptive trends are to be observed.
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References
Atkinson, D.E.: Adaptations of enzymes for regulation of catalytic function. Biochem. Soc. Symp.41, 205–223 (1976)
Borgmann, U., Moon, T.W.: A comparison of lactate dehydrogenases from an ectothermic and an endothermic animal. Canad. J. Biochem.53, 998–1004 (1975)
Baldwin, J.: Adaptation of enzymes to temperature: acetylcholinesterases in the central nervous system of fishes. Comp. Biochem. Physiol.40B, 181–187 (1971)
Cuatrecasas, P.: Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J. biol. Chem.245, 3059–3065 (1970)
DeBurgos, N.M.G., Burgos, C., Gutierrez, M., Blanco, A.: Effect of temperature upon the catalytic properties of lactate dehydrogenase isoenzymes from a poikilotherm. Biochim. biophys. Acta (Amst.)315, 250–258 (1973)
Freed, J.M.: Properties of muscle phosphofructokinase of cold- and warm-acclimatedCarassius auratus. Comp. Biochem. Physiol.39B, 747–764 (1971)
Heisler, N., Weitz, H., Weitz, A.M.: Extracellular and intracellular pH with changes of temperature in the dogfishScyliorhinus stellaris. Resp. Physiol.26, 249–263 (1976)
Hochachka, P.W., Lewis, J.K.: Interacting effects of pH and temperature on theK m values for fish tissue lactate dehydrogenases. Comp. Biochem. Physiol.39B, 925–933 (1971)
Kobayashi, S., Hubbell, H.R., Orengo, A.: A homogeneous, thermostable deoxythymidine kinase fromBacillus stearothermophilus. Biochemistry13, 4537–4543 (1974)
Laidler, K.J., Bunting, P.S.: The chemical kinetics of enzyme action, 2nd ed., pp. 142–162. Oxford: Clarendon Press 1973
Low, P.S., Somero, G.N.: Adaptation of muscle pyruvate kinases to environmental temperatures and pressures. J. exp. Zool.198, 1–12 (1976)
Malan, A., Wilson, T.L., Reeves, R.B.: Intracellular pH in cold-blooded vertebrates as a function of body temperature. Resp. Physiol.28, 29–47 (1976)
O'Carra, P., Barry, S., Corcoran, E.: Affinity chromatographic differentiation of lactate dehydrogenase isoenzymes on the basis of differential abortive complex formation. FEBS Lett.43, 163–168 (1974)
Perrin, D.D., Dempsey, B.: Buffers for pH and metal ion control. New York: Wiley 1974
Rahn, H., Reeves, R.B., Howell, B.J.: Hydrogen ion regulation, temperature, and evolution. Amer. Rev. resp. Dis.112, 165–172 (1975)
Roberts, G.C.K., Meadows, D.H., Jardetzky, O.: Nuclear magnetic resonance studies of the structure and binding sites of enzymes. VII. Solvent and temperature effects on the ionization of histidine residues of ribonuclease. Biochemistry8, 2053–2056 (1969)
Sacks, J., Ganslen, R.V., Diffee, J.T.: Lactic and pyruvic acid relations in frog muscle. Amer. J. Physiol.177, 113–114 (1954)
Spielmann, H., Erickson, R.P., Epstein, C.J.: The separation of lactate dehydrogenase X from other lactate dehydrogenase isozymes of mouse testes by affinity chromatography. FEBS Lett.35, 19–23 (1973)
Vesell, E.S., Pool, P.E.: Lactate and pyruvate concentration in exercised ischemic canine muscle. Relationship of tissue substrate level to lactate dehydrogenase isozyme patterns. Proc. nat. Acad. Sci. (Wash.)55, 756–762 (1966)
Waddell, W.J., Bates, R.G.: Intracellular pH. Physiol. Rev.49, 285–329 (1969)
Wadler, F.C., Hoffmann, F.M.: Glutamine synthetase ofBacillus stearothermophilus. I. Purification and basic properties. Biochemistry13, 3207–3214 (1974)
Wilkinson, G.N.: Statistical estimations in enzyme kinetics. Biochem. J.80, 324–332 (1961)
Wilson, T.L.: Interrelations between pH and temperature for the catalytic rate of the M4 isozyme of lactate dehydrogenase (EC 1.1.1.27) from goldfish (Carassius auratus L.). Arch. Biochem. Biophys.179, 378–390 (1977a)
Wilson, T.L.: Theoretical analysis of the effects of two pH regulation patterns of the temperature sensitivities of biological systems in nonhomeothermic animals. Arch. Biochem. Biophys.182, 409–419 (1977b)
Yancey, P.H., Somero, G.N.: Urea-requiring lactate dehydrogenases of marine elasmobranch fishes. J. comp. Physiol.125, 135–141 (1978)
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Yancey, P.H., Somero, G.N. Temperature dependence of intracellular pH: Its role in the conservation of pyruvate apparentK m values of vertebrate lactate dehydrogenases. J Comp Physiol B 125, 129–134 (1978). https://doi.org/10.1007/BF00686748
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DOI: https://doi.org/10.1007/BF00686748