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Identification and purification of an aspartic proteinase from human semen

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Abstract

To purify and evaluate the molecular changes associated with an aspartic protease (Cathepsin D) in human semen from infertile subjects. Cathepsin D was purified from normo-, oligo- and azoospermic semen, by a procedure involving detergent solubilisation, affinity chromatography and gel filtration chromatography. The enzyme from normo-, oligo- and azoospermic samples was purified 86, 60 and 44 fold respectively. The purified enzyme appeared as a single band on SDS as well as on native PAGE irrespective of the pathological conditions. The molecular weight of Cathepsin D from oligospermic and normospermic samples was 40 kDa while that of azoospermic sample was found to be 43 kDa. The enzyme was inhibited by pepstatin while other proteinase inhibitors and metal ions did not have any effect. Purified Cathepsin D from azoospermic sample differs from normospermia and oligospermia.

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References

  1. Lindquist, F, Thorsteinsson, T. and Baus, O. (1955) Purification and properties of some findings in human seminal fluid. Biochem J. 59, 69–79.

    Google Scholar 

  2. Mann, T. (1964)Biochemistry of semen and the male reproductive tract. London: Methuen & Co. Ltd.

    Google Scholar 

  3. Moghissi, K.S. and Syner, F.N. (1970) Studies on human cervical mucus: Mucoids and their relation to sperm penetration. Fertil Steril. 21 (Suppl 3), 234–239.

    PubMed  CAS  Google Scholar 

  4. Kazuo, Y., Keitaro, M., Masaru, O., Kiyotaka, T., Tomoko, N., Shinichi, K. and Tadash, B. (1988) Acrosine accelerates the dispersal of sperm acrosomal proteins during acrosome reaction. 273 (Suppl 17), 10470–10474.

    Google Scholar 

  5. Daulat, T.R.P., Aida, A.H., Christopher, R.L. and Ben, M.J.P. (1998) The biological and functional significance of the sperm acrosome and acrosomal enzymes in mammalian fertilization. Experimental Cell Research 240 (Suppl 2), 151–164.

    Google Scholar 

  6. Ruenwongsa, P. and Choulavtnatol, M. (1977) Acid protease and its proenzyme from human seminal plasma. Adv Exp Biol. (United States) 95, 329–341.

    CAS  Google Scholar 

  7. Walker, B., Bernstein, G.S., Diedrich, K., Nahamura, R.M. and Kreba, D. (1988) Acrosomal proteinase activity of human spermatozoa and relation of results to semen quality. Human Reprod. 3 (2), 75–80.

    Google Scholar 

  8. Marc, G.L.M., Elisen, Roelof J., van Kooij, Martijn, A., Nolte, J., Arnoud Marquart, Tycho, M.T.W.T., Lock, Bonno, N. Bouma, and Joost C.M. Meijers. (1998) Protein C inhibitor may modulate human sperm-oocyte interaction. Biol. of. Reprod. 58, 670–677.

    Article  Google Scholar 

  9. W.H.O. (1992)Laboratory manual for the examination of human semen and sperm cervical mucus interaction. 3rd Edition, Cambridge University Press, Cambridge.

    Google Scholar 

  10. Miller, G.L. (1959) Protein determination of large number of samples. Anal Chem. 35, 964.

    Article  Google Scholar 

  11. Barrett, A.J. (1977) Cathepsin D and other carboxyl proteases. In:Proteinases in mammalian cells and tissues. Barret, A.J. (ed.). North Holland Publishing Co., Amsterdam, 207.

    Google Scholar 

  12. Corner, G. (1998) Cathepsin D. In:Handbook of proteolytic enzymes. Alan Barrett, Neil Rowlings and J.F. Woesnner (eds.). Acad. Press. 828–836.

  13. Kirschke, H. and Barrett, A.J. (1987) Chemistry of Lysosomal Proteases. In:Lysosomes: Their role in protein breakdown. Gauman, H. and Ballart, F.J. (eds.). Academic Press.

  14. Szecsi, P.B., Halgreen, H., Wong, R.N., Kjaer, T. and Tang, J. (1955) Cellular origin, complementary deoxyribonucleic acid and N-terminal amino acid sequences of human seminal progastricsin. Biol. Reprod. 53 (1), 227–233.

    Article  Google Scholar 

  15. Dandekar, S.P. and Harikumar, P. (1997) Seminal profile of lysosomal enzymes in normal and infertile men. J Postgraduate Med. 43 (ii), 33–37.

    CAS  Google Scholar 

  16. Wassarman, P.M. (1999) Mammalian Fertilization: Molecular aspects of Gamete Adhesion, Exocytosis, and Fusion. Cell 96, 175–183.

    Article  PubMed  CAS  Google Scholar 

  17. Wassarman, P.M., Litscher, E.S. (1998) Towards the molecular basis of sperm and egg interaction during mammalian fertilization. Cell Tissue Organs 2001, 36–45.

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Seth G.S. Medical College & K.E.M. Hospital, Parel, 400 012, Mumbai, India

    S. R. Pardesi & S. P. Dandekar (Prof. & Head)

  2. Food Technology Division, Bhabha Atomic Research Centre, 400 085, Mumbai, India

    S. N. Jamdar & P. Harikumar

Authors
  1. S. R. Pardesi
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  2. S. P. Dandekar
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  3. S. N. Jamdar
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  4. P. Harikumar
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Correspondence to S. P. Dandekar.

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Pardesi, S.R., Dandekar, S.P., Jamdar, S.N. et al. Identification and purification of an aspartic proteinase from human semen. Indian J Clin Biochem 19, 84–90 (2004). https://doi.org/10.1007/BF02894262

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