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Cryptopain-1, a cysteine protease of Cryptosporidium parvum, does not require the pro-domain for folding

  • B.-K. NA (a1), J.-M. KANG (a1), H.-I. CHEUN (a2), S.-H. CHO (a2), S.-U. MOON (a2), T.-S. KIM (a3) and W.-M. SOHN (a1)...


Cryptosporidium parvum is an intracellular protozoan parasite that causes cryptosporidiosis in mammals including humans. In the current study, the gene encoding the cysteine protease of C. parvum (cryptopain-1) was identified and the biochemical properties of the recombinant enzyme were characterized. Cryptopain-1 shared common structural properties with cathepsin L-like papain family enzymes, but lacked a typical signal peptide sequence and contained a possible transmembrane domain near the amino terminus and a unique insert in the front of the mature domain. The recombinant cryptopain-1 expressed in Escherichia coli and refolded to the active form showed typical biochemical properties of cathepsin L-like enzymes. The folding determinant of cryptopain-1 was characterized through multiple constructs with or without different lengths of the pro-domain of the enzyme expressed in E. coli and assessment of their refolding abilities. All constructs, except one that did not contain the full-length mature domain, successfully refolded into the active enzymes, suggesting that cryptopain-1 did not require the pro-domain for folding. Western blot analysis showed that cryptopain-1 was expressed in the sporozoites and the enzyme preferentially degraded proteins, including collagen and fibronectin, but not globular proteins. This suggested a probable role for cryptopain-1 in host cell invasion and/or egression by the parasite.


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*Corresponding author: Department of Parasitology and Institute of Health Sciences,College of Medicine, Gyeongsang National University, Jinju660-751, Korea. Tel: +82 55 751 8822. Fax: +82 55 759 4022. E-mail:


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Cryptopain-1, a cysteine protease of Cryptosporidium parvum, does not require the pro-domain for folding

  • B.-K. NA (a1), J.-M. KANG (a1), H.-I. CHEUN (a2), S.-H. CHO (a2), S.-U. MOON (a2), T.-S. KIM (a3) and W.-M. SOHN (a1)...


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