Home⇛The Philippine Journal of Biochemistry and Molecular Biology (PJBMB)⇛vol. 5 and 6 no. 1 (1982)

Partial Purification and Biochemical Characterization of Rat Sperm Hyaluronidase

Lpurdes Cruz

Discipline: veterinary sciences

 

Abstract:

Hyaluronidase (hyaluronoglucosaminidase; hyaluronate-4-glycanohy- drolase, E.C. 3.2.1.35) from rat cauda epididymal sperm was extracted by the non-ionic detergent, Triton X-100. It had optimum activity at pH 4.0 and 37°C. The unstable nature of the enzyme during storage was observed. Hyaluronidase from rat sperm was partially purified by using DEAEcellulose chromatography, cellulose phosphate chromatography and Sepharose 4B chromatography. Cellulose phosphate column chromatography pro- duced the highest purification and resulted in 232 turbidity reducing units per mg protein. Disc gel electrophoresis showed one major component and a minor band of faster mobility. The Km of hyaluronidase on hyaluronic acid was 0.40 mg per ml, while V was 15.9 mg hyaluronic acid digested per min per mg protein.



References:

  1. Casillas, E. R., & Chulavatnatol, M. (1979). Effect of carnitine on glycolytic intermediate levels and adenosine triphosphate phosphorylation in bovine epididymal spermatozoa. Journal of Reproduction and Fertility, 56(2), 439–444.
  2. Polakoski, K. L., Zaneveld, L. J. D., & Williams, W. L. (1972). Purification of a proteolytic enzyme from rabbit acrosomes. Biology of Reproduction, 6(1), 23–29.
  3. Hartree, E. F., & Srivastava, P. N. (1965). Chemical composition of the acrosomes of ram spermatozoa. Journal of Reproduction and Fertility, 9(1), 47–60.
  4. Toksdorf, S., McCready, M. H., McCullagh, D. R., & Schwenk, E. (1949). The turbidimetric assay of hyaluronidase. Journal of Laboratory and Clinical Medicine, 34(1), 74–89.
  5. Kass, E. H., & Seastone, C. V. (1944). The role of the mucoid polysaccharide (hyaluronic acid) in the virulence of group A hemolytic streptococci. Journal of Experimental Medicine, 79(3), 319–330.
  6. Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265–274.
  7. Whatman. (n.d.). Advanced ion exchange cellulose manual. Whatman Biochemicals Ltd.
  8. Pharmacia Fine Chemicals. (1979). Ion exchange chromatography: Principles and methods (Manual). Rahms i Lund.
  9. Pharmacia Fine Chemicals. (1979). Gel filtration theory and practice (Manual). Rahms i Lund.
  10. Canalco, Inc. (1969). Research disc electrophoresis instruction manual. Canalco, Inc.
  11. McClean, D. (1941). The capsulation of streptococci and its relation to diffusion factor (hyaluronidase). Journal of Pathology and Bacteriology, 53(1), 13–27.
  12. Hoffman, D. C., & Duran-Reynals, F. (1931). The influence of testicle extract on the intradermal injection of homologous and heterologous bacteria. Journal of Experimental Medicine, 53(3), 387–398.
  13. Orgebin-Crist, M. C. (1967). Maturation of spermatozoa in the rabbit epididymis: Fertilizing capacity and embryonic mortality in doe rabbits inseminated with release epididymal spermatozoa. Nature, 217(5131), 816–818.
  14. Cummins, J. M. (1976). Effects of epididymal maturation on the motility and structural longevity of chinchilla spermatozoa in vitro. Journal of Experimental Zoology, 197(2), 187–190.
  15. Zaneveld, L. J. D., Polakoski, K. L., & Schumacher, G. F. B. (1973). Properties of acrosomal hyaluronidase from bull spermatozoa. Journal of Biological Chemistry, 248(2), 564–570.
  16. Yang, C. H., & Srivastava, P. N. (1974). Purification and properties of hyaluronidase from bull spermatozoa acrosomes. Journal of Reproduction and Fertility, 37(1), 17–25.
  17. Borders, C. L., Jr., & Raftery, M. A. (1968). Purification and partial characterization of testicular hyaluronidase. Journal of Biological Chemistry, 243(14), 3756–3762.
  18. Brunish, R., & Högberg, B. (1960). Some of the properties of purified testicular hyaluronidase. Comptes Rendus des Travaux du Laboratoire Carlsberg, 32, 35–47.
  19. Khorlin, A. Y., Vikha, I. V., & Milishnikov, A. N. (1973). Subunit structure of testicular hyaluronidase. FEBS Letters, 31(1), 107–110.
  20. Yang, C. H., & Srivastava, P. N. (1975). Isolation and properties of b-N-acetylglucosaminidase from bull sperm acrosomes. Biochimica et Biophysica Acta (BBA) - Enzymology, 391(2), 382–397.

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