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Open AccessArticle

Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

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State Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai 200237, China
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Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
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School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
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College of Science, Health Education and Engineering, Vector- and Water-Borne Pathogen Research Group, Murdoch University, Murdoch, Western Australia 6150, Australia
*
Authors to whom correspondence should be addressed.
Microorganisms 2019, 7(10), 452; https://doi.org/10.3390/microorganisms7100452
Received: 25 August 2019 / Revised: 7 October 2019 / Accepted: 12 October 2019 / Published: 14 October 2019
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. View Full-Text
Keywords: Cryptosporidium parvum; insulinase-like protease; expression; invasion Cryptosporidium parvum; insulinase-like protease; expression; invasion
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MDPI and ACS Style

Xu, R.; Guo, Y.; Li, N.; Zhang, Q.; Wu, H.; Ryan, U.; Feng, Y.; Xiao, L. Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum. Microorganisms 2019, 7, 452.

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