Skip to main content
SpringerLink
Log in

Partial purification and characterization of acid phosphatase from sporulated oocysts ofEimeria tenella

  • Full Papers
  • Published:
Experientia Aims and scope Submit manuscript

Summary

Acid phosphatase ofEimeria tenella oocysts (Peak II) was purified 77-fold with a recovery of 26% using protamine sulfate precipitation, DEAE-cellulose chromatography and Sephadex G-200 gel filtration. This enzyme occurs in multiple forms as indicated by two peaks which can be separated by DEAE-cellulose chromatography and polyacrylamide gel electrophoresis. The partially purified enzyme has optimal activity at pH 4.5. With p-nitrophenyl phosphate the Km and Vmax values for (Peak II) were 25 mM and 1.57 μmol/min/mg protein, respectively. The enzyme (Peak II) ist strongly inhibited by Hg++, Cu++, iodoacetamide, fluoride and molybdate. Tartrate and other divalent metal ions have no effect on enzyme activity. The partially purified Peak II phosphatase is not a glycoprotein as it is not absorbed on concanavalin-A Sepharose and its treatment with bacterial neuraminidase does not alter its elution profile through DEAE cellulose.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature

  1. Roche, J., Phosphatases, in: Enzymes, vol. 1, pp. 473–510. Eds J. B. Sumner and K. Myrback. Academic Press, New York 1950.

    Google Scholar 

  2. Hollander, V. P., Acid phosphatases, in: Enzymes, vol. 4, pp. 449–497. Ed. P. Boyer. Academic Press, New York 1971.

    Google Scholar 

  3. Gill, B. S., and Ray, H. N., Phosphatases and their significance inEimeria tenella. Ind. J. Vet. Sci. Ani. Husb.24 (1954) 239–244.

    CAS  Google Scholar 

  4. Ray, H. N., and Gill, B. S., Preliminary observations on alkaline phosphatase in experimentalEimeria tenella infections in chicks. Ann. trop. Med. Parasitol48 (1954) 8–10.

    Article  CAS  PubMed  Google Scholar 

  5. Frandsen, J. C.,Eimeria stiedae: cytochemical identification of acid and alkaline phosphatases, carboxylic ester hydrolases and succinate, lactate and glucose 6-phosphate dehydrogenases in endogenous stages or rabbit tissues. Exp. Parasitol.23 (1968) 398–411.

    Article  CAS  PubMed  Google Scholar 

  6. Shirley, M. W., Enzyme variation inEimeria species of chicken. Parasitology71 (1975) 369–376.

    Article  CAS  PubMed  Google Scholar 

  7. Rollinson, D., Electrophoretic variation of enzymes in chicken coccidia. Tran. R. Soc. trop. Med. Hyg.69 (1975) 436–437.

    CAS  Google Scholar 

  8. Farooqui, A. A., Lujan, R., and Hanson, W. L., Acid hydrolases of the coccidian,Eimeria tenella. Experientia39 (1983) 1368–1370.

    Article  CAS  PubMed  Google Scholar 

  9. Vetterling, J. M., Continuous-flow differential density flotation of coccidial oocysts and a comparison with other methods. J. Parasitol.55 (1969) 412–417.

    Article  CAS  PubMed  Google Scholar 

  10. Roberson, E. L., Hanson, W. L., and Chapman Jr, W. L.,Trypanosoma cruzi. Effect of antithymocyte serum in mice and neonatal thymectomy in rats. Exp. Parasitol.34 (1973) 168–180.

    Article  CAS  PubMed  Google Scholar 

  11. Helwig, J. J., Farooqui, A. A., Bollack, C., and Mandel, P., Distribution of lysosomal hydrolases in glomerular and tubular fractions of rabbit kidney cortex. Int. J. Biochem.8 (1977) 323–327.

    Article  CAS  Google Scholar 

  12. Fiske, C. H., and Subba Row, Y., The colorimetric determination of phosphorus. J. biol. Chem.66 (1925) 375–400.

    Article  CAS  Google Scholar 

  13. Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Analyt. Biochem.72 (1976) 248–254.

    Article  CAS  PubMed  Google Scholar 

  14. Brewer, J. M., and Ashworth, R. B., Disc electrophoresis. J. chem. Educ.46 (1969) 41–45.

    Article  CAS  PubMed  Google Scholar 

  15. Farooqui, A. A., Purification and properties of arysulfatase A from human placenta. Archs int. Physiol. Biochem.84 (1976) 479–492.

    CAS  Google Scholar 

  16. Andrews, P., The gel filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J.96 (1965) 595–606.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ryley, J. F., Bentley, M., Manners, D. J., and Stark, J. R., Amylopectin, the storage polysaccharide of the coccidiaEimeria brunetti andE. tenella. J. Parasitol.55 (1969) 839–845.

    Article  CAS  PubMed  Google Scholar 

  18. Helwig, J. J., Farooqui, A. A., Bollack, C., and Mandel, P., Purification and some properties of tartrate-sensitive acid phosphatase from rabbit kidney cortex. Biochem. J.175 (1978) 321–329.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Farooqui, A. A. and Hanson, W. L., Comparison of arylsulphatases fromEimeria tenella (parasite) and chicken caecum (host). Biochem. J.242 (1987) 97–102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Patel, A., and Koenig, H., Brain lysosomal hydrolases I. Solubilization and electrophoretic behavior of acid hydrolases in nerve-ending and mitochondrial-lysosomal fractions from rat brain. Effect of autolysis, neuraminidase and storage. Neurochem. Res.1 (1976) 275–298.

    Article  CAS  PubMed  Google Scholar 

  21. Farooqui, A. A., Adam, D. D., Hanson, W. L., and Prestwood, A. K., Studies on the enzymes ofSarcocystis suicanis: Purification and characterization of an acid phosphatase. J. Parasitol.73 (1987) 681–688.

    Article  CAS  PubMed  Google Scholar 

  22. Lam, K., Lai, L., Burkart, P. T., and Yam, L. T., Kinetic properties of tartrate resistant acid phosphatase isolated from human spleen with leukemic reticuloendotheliosis. J. biol. Chem.252 (1977) 3371–3373.

    Article  CAS  PubMed  Google Scholar 

  23. Krebs, E. G., and Beavo, J. A., Phosphorylation-dephosphorylation of enzymes. A. Rev. Biochem.48 (1979) 923–959.

    Article  CAS  Google Scholar 

  24. Cohen, P., The role of protein phosphorylation in neural and hormonal control of cellular activity. Nature296 (1982) 613–620.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Parasitology, College of Veterinary Medicine, The University of Georgia, 30602, Athens, Georgia, USA

    A. A. Farooqui & W. L. Hanson

Authors
  1. A. A. Farooqui
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W. L. Hanson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Farooqui, A.A., Hanson, W.L. Partial purification and characterization of acid phosphatase from sporulated oocysts ofEimeria tenella. Experientia 44, 437–440 (1988). https://doi.org/10.1007/BF01940540

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01940540

Key words

Search

Navigation