Spontaneous Proteolytic Processing of Human Recombinant Anti-Mullerian Hormone: Structural and Functional Differences of the Molecular Forms


The technology for the production of highly purified human recombinant anti-mullerian hormone (AMH)—a potential antitumor agent for the treatment of certain types of malignant neoplasms—is described. It was found that spontaneous proteolytic processing of the hormone is possible during the storage of AMH preparations under physiological conditions. This leads to the formation of C-terminal homodimer of AMH (activated form) and, later, to an inactive state during the further proteolysis. Sites at which spontaneous processing of the hormone molecule occurred during prolonged storage with the formation of active and inactive fragments were identified. The structural and functional differences in the molecular forms of the C-terminal fragment contained in the preparations are analyzed.

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

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.
Fig. 7.
Fig. 8.
Fig. 9.
Fig. 10.
Fig. 11.
Fig. 12.


  1. 1

    Gukasova, N.V. and Severin, S.E., Vopr. Biol. Med. Farm. Khim., 2005, no. 4, pp. 3–9.

  2. 2

    Donahoe, P.K., Clarke, T., Teixeira, J., Maheswaran, S., and MacLaughlin, D.T., Mol. Cell. Endocrinol., 2003, vol. 211, no. 1, pp. 37–42.

    Article  CAS  PubMed  Google Scholar 

  3. 3

    Jung, Y.S., Kim, H.J., Seo, S.K., Choi, Y.S., Nam, E.J., Kim, S.W., Han, H.D., Kim, J.W., and Kim, Y.T., Yonsei Med. J., 2016, vol. 57, no. 1, pp. 33–40.

    Article  CAS  PubMed  Google Scholar 

  4. 4

    Pankhurst, M.W., Leathart, B.L., Batchelor, N.J., and McLennan, I.S., Endocrinology, 2016, vol. 157, no. 4, pp. 1622–1629.

    Article  CAS  PubMed  Google Scholar 

  5. 5

    Di Clemente, N., Jamin, S.P., Lugovskoy, A., Carmillo, P., Ehrenfels, C., Picard, J.Y., Whitty, A., Josso, N., Pepinsky, R.B., and Cate, R.L., Mol. Endocrinol., 2010, vol. 24, no. 11, pp. 2193–2206.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. 6

    Nachtigal, M.W. and Ingraham, H.A., Proc. Natl. Acad. Sci. U. S. A., 1996, vol. 93, no. 15, pp. 7711–7716.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    Ragin, R.C., Donahoe, P.K., Kenneally, M.K., Ahmad, M.F., and MacLaughlin, D.T., Protein Expr. Purif., 1992, vol. 3, no. 3, pp. 236–245.

    Article  CAS  PubMed  Google Scholar 

  8. 8

    Rak, A.Ya., Trofimov, A.V., Protasov, E.A., Simbirtsev, A.S., and Ishchenko, A.M., Ross. Immunol. Zh., 2017, vol. 11 (20), no. 4, pp. 755–757.

  9. 9

    Pepinsky, R.B., Sinclair, L.K., Chow, E.P., Mattaliano, R.J., Manganaro, T.F., Donahoe, P.K., and Cate, R.L., J. Biol. Chem., 1988, vol. 263, no. 35, pp. 18961–18964.

    CAS  PubMed  Google Scholar 

  10. 10

    Walker, J.M., Methods Mol. Biol., 1984, vol. 1, pp. 57–61.

    CAS  PubMed  Google Scholar 

  11. 11

    Lorenzo, H.K., Teixeira, J., Pahlavan, N., Laurich, V.M., Donahoe, P.K., and MacLaughlin, D.T., J. Chromatogr. B, 2002, vol. 766, no. 1, pp. 89–98.

    Article  CAS  Google Scholar 

  12. 12

    Massagué, J., Annu. Rev. Biochem., 1998, vol. 67, no. 1, pp. 753–791.

    Article  PubMed  Google Scholar 

Download references


The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

Author information



Corresponding author

Correspondence to A. Ya. Rak.

Additional information

Translated by P. Kuchina

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Rak, A.Y., Trofimov, A.V., Protasov, E.A. et al. Spontaneous Proteolytic Processing of Human Recombinant Anti-Mullerian Hormone: Structural and Functional Differences of the Molecular Forms. Appl Biochem Microbiol 55, 13–20 (2019). https://doi.org/10.1134/S0003683819010149

Download citation


  • anti-mullerian hormone
  • AMH
  • chromatography
  • MALDI mass-spectrometry
  • monoclonal antibodies
  • recombinant protein
  • proteolysis