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

R. Xu; Y. Guo; N. Li; Q. Zhang; H. Wu; U. Ryan; Y. Feng; L. Xiao

doi:10.3390/microorganisms7100452

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

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

R. Xu, Y. Guo, N. Li, Q. Zhang, H. Wu, U. Ryan, Y. Feng and L. Xiao

Microorganisms, Vol.7(10)

2019

DOI: https://doi.org/10.3390/microorganisms7100452

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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.

Details

Title: Characterization of INS-15, a metalloprotease potentially involved in the invasion of Cryptosporidium parvum
Authors/Creators: R. Xu (Author/Creator) - East China University of Science and Technology
Y. Guo (Author/Creator) - South China Agricultural University
N. Li (Author/Creator) - South China Agricultural University
Q. Zhang (Author/Creator) - East China University of Science and Technology
H. Wu (Author/Creator) - East China University of Science and Technology
U. Ryan (Author/Creator) - Murdoch University
Y. Feng (Author/Creator) - South China Agricultural University
L. Xiao (Author/Creator) - South China Agricultural University
Publication Details: Microorganisms, Vol.7(10)
Publisher: MDPI
Identifiers: 991005542939807891
Copyright: © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
Murdoch Affiliation: College of Science, Health, Engineering and Education
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.246 Diarrheal Diseases; 1.246.985 Cryptosporidium
Web Of Science research areas: Microbiology
ESI research areas: Microbiology