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

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...

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Autores principales: Xu, Rui, Guo, Yaqiong, Li, Na, Zhang, Qiang, Wu, Haizhen, Ryan, Una, Feng, Yaoyu, Xiao, Lihua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6843835/
https://www.ncbi.nlm.nih.gov/pubmed/31615118
http://dx.doi.org/10.3390/microorganisms7100452
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author Xu, Rui
Guo, Yaqiong
Li, Na
Zhang, Qiang
Wu, Haizhen
Ryan, Una
Feng, Yaoyu
Xiao, Lihua
author_facet Xu, Rui
Guo, Yaqiong
Li, Na
Zhang, Qiang
Wu, Haizhen
Ryan, Una
Feng, Yaoyu
Xiao, Lihua
author_sort Xu, Rui
collection PubMed
description 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.
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spelling pubmed-68438352019-11-25 Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum Xu, Rui Guo, Yaqiong Li, Na Zhang, Qiang Wu, Haizhen Ryan, Una Feng, Yaoyu Xiao, Lihua Microorganisms Article 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. MDPI 2019-10-14 /pmc/articles/PMC6843835/ /pubmed/31615118 http://dx.doi.org/10.3390/microorganisms7100452 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xu, Rui
Guo, Yaqiong
Li, Na
Zhang, Qiang
Wu, Haizhen
Ryan, Una
Feng, Yaoyu
Xiao, Lihua
Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title_full Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title_fullStr Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title_full_unstemmed Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title_short Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum
title_sort characterization of ins-15, a metalloprotease potentially involved in the invasion of cryptosporidium parvum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6843835/
https://www.ncbi.nlm.nih.gov/pubmed/31615118
http://dx.doi.org/10.3390/microorganisms7100452
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