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Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum
Cryptosporidium parvum is a major cause of moderate-to-severe diarrhea in humans and animals. Its compact genome contains 22 genes encoding divergent insulinase-like proteases (INS), which are poorly characterized. In this study, two small members of this family, INS-21 encoded by cgd7_2080 and INS-...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8175991/ https://www.ncbi.nlm.nih.gov/pubmed/34093467 http://dx.doi.org/10.3389/fmicb.2021.651512 |
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author | Xu, Rui Lai, Cong Yang, Fuxian Zhang, Qiang Li, Na Guo, Yaqiong Xiao, Lihua Feng, Yaoyu |
author_facet | Xu, Rui Lai, Cong Yang, Fuxian Zhang, Qiang Li, Na Guo, Yaqiong Xiao, Lihua Feng, Yaoyu |
author_sort | Xu, Rui |
collection | PubMed |
description | Cryptosporidium parvum is a major cause of moderate-to-severe diarrhea in humans and animals. Its compact genome contains 22 genes encoding divergent insulinase-like proteases (INS), which are poorly characterized. In this study, two small members of this family, INS-21 encoded by cgd7_2080 and INS-23 encoded by cgd5_3400, were cloned, expressed, and characterized to understand their functions. Recombinant INS-21 and INS-23 were expressed in Escherichia coli and polyclonal antibodies against these two proteins were prepared. The cgd7_2080 gene had a high transcription level during 0–2 h of in vitro C. parvum culture, while cgd5_3400 was highly transcribed at 0–6 h. INS-21 was mostly located in the apical region of sporozoites and merozoites whereas INS-23 was found as spots in sporozoites and merozoites. The immunoelectron microscopy confirmed the expression of INS-21 in the apical region of sporozoites while INS-23 appeared to be expressed in the dense granules of sporozoites. The neutralization efficiency was approximately 35%, when the cultures were treated with anti-INS23 antibodies. These results suggest that INS-21 and INS-23 are expressed in different organelles and might have different functions in the development of C. parvum. |
format | Online Article Text |
id | pubmed-8175991 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-81759912021-06-05 Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum Xu, Rui Lai, Cong Yang, Fuxian Zhang, Qiang Li, Na Guo, Yaqiong Xiao, Lihua Feng, Yaoyu Front Microbiol Microbiology Cryptosporidium parvum is a major cause of moderate-to-severe diarrhea in humans and animals. Its compact genome contains 22 genes encoding divergent insulinase-like proteases (INS), which are poorly characterized. In this study, two small members of this family, INS-21 encoded by cgd7_2080 and INS-23 encoded by cgd5_3400, were cloned, expressed, and characterized to understand their functions. Recombinant INS-21 and INS-23 were expressed in Escherichia coli and polyclonal antibodies against these two proteins were prepared. The cgd7_2080 gene had a high transcription level during 0–2 h of in vitro C. parvum culture, while cgd5_3400 was highly transcribed at 0–6 h. INS-21 was mostly located in the apical region of sporozoites and merozoites whereas INS-23 was found as spots in sporozoites and merozoites. The immunoelectron microscopy confirmed the expression of INS-21 in the apical region of sporozoites while INS-23 appeared to be expressed in the dense granules of sporozoites. The neutralization efficiency was approximately 35%, when the cultures were treated with anti-INS23 antibodies. These results suggest that INS-21 and INS-23 are expressed in different organelles and might have different functions in the development of C. parvum. Frontiers Media S.A. 2021-05-21 /pmc/articles/PMC8175991/ /pubmed/34093467 http://dx.doi.org/10.3389/fmicb.2021.651512 Text en Copyright © 2021 Xu, Lai, Yang, Zhang, Li, Guo, Xiao and Feng. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Xu, Rui Lai, Cong Yang, Fuxian Zhang, Qiang Li, Na Guo, Yaqiong Xiao, Lihua Feng, Yaoyu Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title | Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title_full | Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title_fullStr | Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title_full_unstemmed | Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title_short | Preliminary Characterization of Two Small Insulinase-Like Proteases in Cryptosporidium parvum |
title_sort | preliminary characterization of two small insulinase-like proteases in cryptosporidium parvum |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8175991/ https://www.ncbi.nlm.nih.gov/pubmed/34093467 http://dx.doi.org/10.3389/fmicb.2021.651512 |
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