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Cryptostatin, a chagasin-family cysteine protease inhibitor of Cryptosporidium parvum

  • J.-M. KANG (a1), H.-L. JU (a1), J.-R. YU (a2), W.-M. SOHN (a1) and B.-K. NA (a1)...

Summary

Cysteine proteases of pathogenic protozoan parasites play pivotal roles in the life cycle of parasites, but strict regulation of their activities is also essential for maintenance of parasite physiology and interaction with hosts. In this study, we identified and characterized cryptostatin, a novel inhibitor of cysteine protease (ICP) of Cryptosporidium parvum. Cryptostatin showed low sequence identity to other chagasin-family ICPs, but 3 motifs (NPTTG, GXGG, and RPW/F motifs), which are evolutionarily conserved in chagasin-family ICPs, were found in the sequence. The overall structure of cryptostatin consisted of 8 β-strands that progressed in parallel and closely resembled the immunoglobulin fold. Recombinant cryptostatin inhibited various cysteine proteases, including papain, human cathepsin B, human cathepsin L, and cryptopain-1, with Ki's in the picomolar range. Cryptostatin was active over a wide pH range and was highly stable under physiological conditions. The protein was thermostable and retained its inhibitory activity even after incubation at 95°C. Cryptostatin formed tight complexes with cysteine proteases, so the complexes remained intact in the presence of sodium dodecyl sulfate and β-mercaptoethanol, but they were disassembled by boiling. An immunogold electron microscopy analysis demonstrated diffused localization of cryptostatin within oocystes and meronts, but not within trophozoites, which suggests a possible role for cryptostatin in host cell invasion by C. parvum.

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Corresponding author

*Corresponding author: Department of Parasitology and Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 660–751, Korea. Tel: +82 55 772 8102. Fax: +82 55 772 8109. E-mail address: bkna@gnu.ac.kr

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Keywords

Cryptostatin, a chagasin-family cysteine protease inhibitor of Cryptosporidium parvum

  • J.-M. KANG (a1), H.-L. JU (a1), J.-R. YU (a2), W.-M. SOHN (a1) and B.-K. NA (a1)...

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