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Characterization of INS-15, a metalloprotease potentially involved in the invasion of Cryptosporidium parvum

Xu, R., Guo, Y., Li, N., Zhang, Q., Wu, H., Ryan, U.ORCID: 0000-0003-2710-9324, Feng, Y. and Xiao, L. (2019) Characterization of INS-15, a metalloprotease potentially involved in the invasion of Cryptosporidium parvum. Microorganisms, 7 (10). p. 452.

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Abstract

Cryptosporidium parvum is a protozoan parasite that can cause moderate-to-severe diarrhea. Insulinase-like proteases (INS) are one of the largest protein families within the small proteome of the pathogen. However, their roles in C. parvum biology remain un-elucidated. In this study, a member of the protein family, INS-15 of C. parvum encoded by cgd3_4260, was cloned, expressed and characterized to understand its function. INS-15 and its domain I were expressed in Escherichia coli and polyclonal antibodies against the domain I and one specific polypeptide were prepared in rabbits. The role of INS-15 protein in the C. parvum invasion was preliminarily studied. Recombinant INS-15 protein and its domain I were successfully expressed in E. coli, together with various degraded products. The cgd3_4260 gene had a peak expression at 2 h of in vitro C. parvum culture, while the INS-15 protein was expressed in the mid-anterior region of sporozoites and the area of merozoites opposite to the nucleus. Anti-INS-15 domain I antibodies reduced the invasion of C. parvum sporozoites by over 40%. The anterior location of INS-15 in invasion stages and partial reduction of in vitro growth indicate that INS-15 plays some roles in the invasion or early development of C. parvum.

Item Type: Journal Article
Murdoch Affiliation: College of Science, Health, Engineering and Education
Publisher: MDPI
Copyright: © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
URI: http://researchrepository.murdoch.edu.au/id/eprint/52429
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