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Partial purification and characterization of a soluble acid phosphatase from the tapeworm, Hymenolepis diminuta

Published online by Cambridge University Press:  05 June 2009

Michael J. Bumbulis  and
Peter W. Pappas
Michael J. Bumbulis
Affiliation:
Department of Zoology, The Ohio State University, Columbus, OH 43120–1293, USA
Peter W. Pappas*
Affiliation:
Department of Zoology, The Ohio State University, Columbus, OH 43120–1293, USA
*
*To whom correspondence should he addressed
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Abstract

An acid phosphatase activity (APA; EC 3.1.3.2) was demonstrated in homogenates of adult Hymenolepis diminuta. The APA was soluble based on the observation that it did not sediment at 130 000 g. APA was partially purified using a combination of differential centrifugation, ammonium sulphate precipitation, chloroform extraction, and gel and fast-protein-liquid-chromatography. This combination of techniques resulted in a preparation with a specific activity approximately 500 times greater than the crude enzyme preparation. The temperature and pH optima of the partially purified APA were 44°C and pH 5·0. The enzyme appeared to be a monomer with a molecular weight of approximately 62 000. APA had a higher affinity for a greater activity towards aromatic than aliphatic phosphoesters and phosphoryl transferase activity was demonstrable using 1-butanol and ethylene glycol as acceptors. APA was inhibited significantly by sodium dodecyl sulphate, fluoride, molybdate and tartrate, but CuSO4 and Fast Garnet GBC were poor inhibitors. The precise cellular localization and function of this enzyme remains unknown since it possesses characteristics of both cytoplasmic and lysosomal APA's of other organisms.

Keywords

Hymenolepis diminutaCestodaacid phosphatasephosphoryl transferasetartratefluoride
Type
Research Papers
Information
Journal of Helminthology , Volume 65 , Issue 2 , June 1991 , pp. 103 - 110
Copyright
Copyright © Cambridge University Press 1991

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References

REFERENCES

Barrett, J. (1981) Biochemistry of Parasitic Helminths. 308 pp. University Park Press: Baltimore.CrossRefGoogle Scholar
Bradford, M. M. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.CrossRefGoogle Scholar
von Brand, T. (1979) Biochemistry and Physiology of Endoparasites. 447 pp. Elsevier/North Holland Biomedical Press: New York.Google Scholar
Chen, C.-H. & Chen, S. C. (1988) Evidence of acid phosphatase in the cytoplasm as a distinct entity. Archives of Biochemistry and Biophysics, 262, 427–438.CrossRefGoogle ScholarPubMed
Fernley, H. N. (1971) Mammalian alkaline phosphatases. In: The Enzymes. Vol. IV. 3rd ed. (editor, Boyer, P. D.) pp. 417–447. Academic Press, New York.Google Scholar
Fioravanti, C. F. & Saz, H. J. (1980) Energy metabolism of adult Hymenolepis diminuta. In: Biology of the Tapeworm Hymenolepis diminuta (editor, Arai, H. P.) pp. 463–504. Academic Press: New York.CrossRefGoogle Scholar
Halton, D. W. (1967) Studies on phosphatase activity in Trematoda. Journal of Parasitology, 53, 46–54.CrossRefGoogle ScholarPubMed
Hollander, V. P. (1971) Acid phosphatases. In: The Enzymes, Vol. IV, 3rd ed. (editor, Boyer, P. D.) pp. 449–498. Academic Press: New York.Google Scholar
Laemmli, U. K. (1970) Cleavage of structural proteins during assembly of bacteriophage T4. Nature, 227, 680–685.CrossRefGoogle ScholarPubMed
Lanzetta, P. A., Alvarez, L. J., Reinach, P. S. & Candia, O. A. (1979) An improved assay for nanomole amounts of inorganic phosphate. Analytical Biochemistry, 100, 95–97.CrossRefGoogle ScholarPubMed
Leis, J. F. & Kaplan, N. O. (1982) An acid phosphatase in the plasma membrane of human astrocytoma showing marked specificity toward phosphotyrosine protein. Proceedings of the National Academy of Sciences of the United States of America, 79, 6507–6511.CrossRefGoogle ScholarPubMed
Li, H-C., Chernoff, J., Chen, L. B. & Kirschobaum, A. (1984) A phosphotyrosyl-protein phosphatase activity associated with acid phosphatase from human prostate gland. European Journal of Biochemistry, 138, 45–51.CrossRefGoogle ScholarPubMed
Lin, M.-H. & Clinton, G. (1986) Human prostatic acid phosphatase has phosphotyrosyl protein phosphatase activity. Biochemical Journal, 235, 351–357.CrossRefGoogle ScholarPubMed
Nigram, V. N. & Fishman, W. H. (1959) Catalysis of phosphoryl transfer by prostatic acid phosphatase. Journal of Biological Chemistry, 234, 2394–2398.CrossRefGoogle Scholar
Panara, F. (1985) Isolation and partial characterization of high and low molecular weight acid phosphatases from chicken liver. International Journal of Biochemistry, 17, 1213–1217.CrossRefGoogle ScholarPubMed
Pappas, P. W. (1982) Hymenolepis diminuta: partial characterization of the membrane-bound and solubilized alkaline phosphohydrolase activities of the isolated brush border plasma membrane. Experimental Parasitology, 54, 80–86.CrossRefGoogle ScholarPubMed
Pappas, P. W. (1988) Acid phosphatase activity in the isolated brush border membrane of the tapeworm, Hymenolepis diminuta: partial characterization and differentiation from the alkaline phosphatase activity. Journal of Cellular Biochemistry, 37, 395–403.CrossRefGoogle ScholarPubMed
Pappas, P. W. & Leiby, D. A. (1986) Variation in the sizes of eggs and oncosphere and the numbers and distributions of testes in the tapeworm, Hymenolepis diminuta. Journal of Parasitology, 72, 383–391.CrossRefGoogle ScholarPubMed
Read, C. P., Rothman, A. H. & Simmons, J. E. Jr (1963) Studies on membrane transport, with special reference to parasite-host integration. Annals of the New York Academy of Sciences, 113, 154–205.CrossRefGoogle ScholarPubMed
Sensabaugh, G. F. (1975) Genetic and non-genetic variation of human acid phosphatases. In: Isozymes, Vol. 1 (editor, Markert, C. L.). pp. 367–380. Academic Press: New York.CrossRefGoogle Scholar
Smyth, J. D. & Mcmanus, D. P. (1989) The Physiology and Biochemistry of Cestodes, 398 pp. Cambridge University Press: Cambridge.CrossRefGoogle Scholar
Uglem, G. L. & Just, J. J. (1983) Trypsin inhibition by tapeworms: antienzyme secretion or pH adjustment? Science, 220, 79–81.CrossRefGoogle ScholarPubMed