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Biochemistry (Moscow)

, Volume 75, Issue 7, pp 866–872 | Cite as

Isolation, purification, and study of properties of recombinant hepsin from Escherichia coli

  • A. A. RaevskayaEmail author
  • E. M. Kuznetsova
  • M. V. Savvateeva
  • S. E. Severin
Article

Abstract

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.

Key words

hepsin proteolytic activity anthralin inhibition cytotoxicity human prostate adenocarcinoma 

Abbreviations

HAI-1 (2)

hepatocyte growth factor activator inhibitor type I (II)

IPTG

isopropyl-1-thio-β-D-galactopyranoside

P1, P2, P3 and P4

are the specific cleavage site residues enumerated from the C- to N-terminus of the peptide

PSA

prostate-specific antigen

SRCR-domain

scavenger cysteine-rich receptor domain

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Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  • A. A. Raevskaya
    • 1
    Email author
  • E. M. Kuznetsova
    • 2
  • M. V. Savvateeva
    • 1
  • S. E. Severin
    • 2
  1. 1.Biological FacultyLomonosov Moscow State UniversityMoscowRussia
  2. 2.Moscow Research Institute of Medical EcologyMoscowRussia

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