Summary
Microsomes were prepared from perfused rat livers after different perfusion procedures. The yield of microsomal protein and the kinetic data (K m, V max) of glucose-6-phosphatase (3.1.3.9) and esterase (3.1.1.1) activities were analysed in each preparation.
No marked differences were detected between conventionally prepared liver microsomes and those from livers perfused 1 hr with an erythrocytes-free medium under the conditions of open outflow. If the outflow pressure was increased artificially, the yield of microsomal protein decreased. The V max of both enzymes was markedly increased, whereas the K m values remained unchanged. The same microsomal alterations occurred when perfused rat livers were poisoned with phalloidin in vitro under the condition of open outflow.
Our findings indicate that microsomal alterations in livers from poisoned animals might be due to microcirculatory disturbances, and not primary effects of the toxin on the endoplasmatic reticulum.
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References
Ames, B. W., Dubin, D. T.: The role of polyamines in the neutralization of bacteriophage desoxyribonucleic acid. J. biol. Chem. 235, 769–775 (1960)
Arion, W. J., Wallin, K. B., Carlson, P. W., Lange, A. J.: The specificity of glucose-6-phosphatase of intact liver microsomes. J. biol. Chem. 247, 2558–2565 (1972)
Bode, C., Goebell, H., Stabler, H.: Zur Eliminierung von Trübungsfehlern bei der Eiweißbestimmung mit der Biuret-Methode. Z. klin. Chem. klin. Biochem. 6, 418–422 (1968)
Degkwitz, E., Köhler, F. R., Staudinger, Hj.: Aktivitätsänderungen von einigen mikrosomalen Leberenzymen nach Phalloidinvergiftung bei Ratten. Hoppe-Seyler's Z. physiol. Chem. 350, 1023–1026 (1969)
Frimmer, M., Gries, J., Hegner, D., Schnorr, B.: Studies on the mechanism of the action of phalloidin. Release of lysosomal enzymes and potassium. Naunyn-Schmiedebergs Arch. Pharmak. exp. Path. 258, 197–214 (1967)
Frimmer, M.: The influence of physical conditions on swelling and K+-release in perfused rat livers poisoned with phalloidin. Naunyn-Schmiedeberg's Arch. Pharmacol. 275, 393–403 (1972)
Frimmer, M., Kroker, R., Porstendörfer, J.: The mode of action of phalloidin: Demonstration of rapid deformation of isolated hepatocytes by scanning electron microscopy. Naunyn-Schmiedeberg's Arch. Pharmacol. 284, 395–398 (1974)
Harper, A. E.: Glucose-6-phosphatase. In: Methoden der enzymatischen Analyse, ed. by H. U. Bergmeyer, pp. 782–792. Weinheim (Bergstr.): Verlag Chemie 1962
Hogeboom, G. H., Schneider, W. C., Palade, G.: Cytochemical studies of mammalian tissues. J. biol. Chem. 172, 619–635 (1948)
Jahn, W.: Similarity between the effect of experimental congestion of the isolated perfused rat livers and the action of cytochalasin B. Naunyn-Schmiedeberg's Arch. Pharmacol. 278, 431–434 (1973)
Krisch, K.: Eigenschaften und Substratspezifität einer Esterase aus Schweinelebermikrosomen. Biochem. Z. 337, 546–573 (1973)
Miller, F., Wieland, O.: Elektronenmikroskopische Untersuchungen zum Wirkungsmechanismus des Knollenblätterpilzgiftes Phalloidin. Virchows Arch. path. Anat. 343, 83–99 (1967)
Nordlie, R. C., Arion, W., Hanson, T. L., Gilsdorf, J. R., Horne, R. N.: Biological regulation of liver microsomal inorganic pyrophosphate-glucose phosphotransferase, glucose-6-phosphatase and inorganic pyrophosphatase. J. biol. Chem. 243, 1140–1146 (1968)
Rick, W.: Chymotrypsin. In: Methoden der enzymatischen Analyse, ed. H. U. Bergmeyer, pp. 800–806. Weinheim (Bergstr.): Verlag Chemie 1962
Stetten, M. R., Burnett, F. F.: Activation of rat-liver microsomal glucose-6-phosphatase inorganic pyrophosphatase, and inorganic pyrophosphate-glucose phosphotransferase by hydroxyl ion. Biochim. biophys. Acta (Amst.) 128, 344–350 (1966)
Tuchweber, B., Kovacs, K., Khandekar, J. D., Garg, B. D.: Peliosis-like changes induced by phalloidin in the rat liver. A light and electron microscopic study. J. Med. 4, 327–345 (1973)
Schneider, W. C.: Intracellular distribution of enzymes. J. biol. Chem. 176, 259–266 (1948)
Wang, E.: The effect of cytochalasin B on endocytosis in BHK-21 cells. J. Cell. Biol. 55, 273 a (1972)
Weiss, E., Sterz, I., Frimmer, M., Kroker, R.: Electron microscopy of isolated rat hepatocytes, before and after treatment with phalloidin. Beitr. Path. 150, 345–356 (1973)
Wieland, O.: Changes in liver metabolism induced by the poisons of amanita phalloides. Clin. Chem.2, 323–338 (1965)
Zurier, R. B., Hoffstein, S., Weissmann, G.: Cytochalasin B: Effect of lysosomal enzyme release from human leucocytes. Proc. nat. Acad. Sci. (Wash.) 70, 844–848 (1973)
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This work was supported by the Deutsche Forschungsgemeinschaft.
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Homann, J., Frimmer, M. Glucose-6-phosphatase (EC 3.1.3.9) and esterase (EC 3.1.1.1) activities of microsomes prepared from perfused rat livers after partial outflow block or phalloidin poisoning. Naunyn-Schmiedeberg's Arch. Pharmacol. 288, 87–96 (1975). https://doi.org/10.1007/BF00501816
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DOI: https://doi.org/10.1007/BF00501816