Isolation, Purification, and Study of Properties of Recombinant Hepsin from Escherichia coli

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A. A. Raevskaya^1*, E. M. Kuznetsova², M. V. Savvateeva¹, and S. E. Severin²

¹Biological Faculty, Lomonosov Moscow State University, 119992 Moscow, Russia; E-mail: araevskaia@gmail.com

²Moscow Research Institute of Medical Ecology, Simferopolskii Bulvar 8, 117638 Moscow, Russia; E-mail: sergsev@aha.ru

^* To whom correspondence should be addressed.

Received November 13, 2009; Revision received December 8, 2009

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A recombinant hepsin-producing strain of Escherichia coli was obtained and the conditions for hepsin expression in a bacterial system were optimized. To study the physicochemical properties of the enzyme, a procedure for purification of active recombinant hepsin using metal-chelate affinity chromatography and ion-exchange chromatography was developed. The interaction of recombinant hepsin with various peptide substrates is characterized. The dose-dependent inhibition of the recombinant hepsin enzyme activity by anthralin in vitro and an increase in the hepsin enzymatic activity in the presence of resveratrol were revealed.

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KEY WORDS: hepsin, proteolytic activity, anthralin, inhibition, cytotoxicity, human prostate adenocarcinoma

DOI: 10.1134/S0006297910070084

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