Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1783468309093548032 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6843835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT xurui characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT guoyaqiong characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT lina characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT zhangqiang characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT wuhaizhen characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT ryanuna characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT fengyaoyu characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum AT xiaolihua characterizationofins15ametalloproteasepotentiallyinvolvedintheinvasionofcryptosporidiumparvum |