Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40
Abstract Background Cryptosporidium parvum is an apicomplexan zoonotic parasite causing the diarrheal illness cryptosporidiosis in humans and animals. To invade the host intestinal epithelial cells, parasitic proteins expressed on the surface of sporozoites interact with host cells to facilitate the...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13071-024-06233-5 |
| _version_ | 1797247509987328000 |
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| author | Yuexin Wang Na Li Guanda Liang Luyang Wang Xiaotian Zhang Zhaohui Cui Xiaoying Li Sumei Zhang Longxian Zhang |
| author_facet | Yuexin Wang Na Li Guanda Liang Luyang Wang Xiaotian Zhang Zhaohui Cui Xiaoying Li Sumei Zhang Longxian Zhang |
| author_sort | Yuexin Wang |
| collection | DOAJ |
| description | Abstract Background Cryptosporidium parvum is an apicomplexan zoonotic parasite causing the diarrheal illness cryptosporidiosis in humans and animals. To invade the host intestinal epithelial cells, parasitic proteins expressed on the surface of sporozoites interact with host cells to facilitate the formation of parasitophorous vacuole for the parasite to reside and develop. The gp40 of C. parvum, named Cpgp40 and located on the surface of sporozoites, was proven to participate in the process of host cell invasion. Methods We utilized the purified Cpgp40 as a bait to obtain host cell proteins interacting with Cpgp40 through the glutathione S-transferase (GST) pull-down method. In vitro analysis, through bimolecular fluorescence complementation assay (BiFC) and coimmunoprecipitation (Co-IP), confirmed the solid interaction between Cpgp40 and ENO1. In addition, by using protein mutation and parasite infection rate analysis, it was demonstrated that ENO1 plays an important role in the C. parvum invasion of HCT-8 cells. Results To illustrate the functional activity of Cpgp40 interacting with host cells, we identified the alpha-enolase protein (ENO1) from HCT-8 cells, which showed direct interaction with Cpgp40. The mRNA level of ENO1 gene was significantly decreased at 3 and 24 h after C. parvum infection. Antibodies and siRNA specific to ENO1 showed the ability to neutralize C. parvum infection in vitro, which indicated the participation of ENO1 during the parasite invasion of HCT-8 cells. In addition, we further demonstrated that ENO1 protein was involved in the regulation of cytoplasmic matrix of HCT-8 cells during C. parvum invasion. Functional study of the protein mutation illustrated that ENO1 was also required for the endogenous development of C. parvum. Conclusions In this study, we utilized the purified Cpgp40 as a bait to obtain host cell proteins ENO1 interacting with Cpgp40. Functional studies illustrated that the host cell protein ENO1 was involved in the regulation of tight junction and adherent junction proteins during C. parvum invasion and was required for endogenous development of C. parvum. Graphical Abstract |
| first_indexed | 2024-04-24T19:59:50Z |
| format | Article |
| id | doaj.art-fc68176b72b0490db64a691d3b50208b |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-04-24T19:59:50Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-fc68176b72b0490db64a691d3b50208b2024-03-24T12:12:00ZengBMCParasites & Vectors1756-33052024-03-0117111310.1186/s13071-024-06233-5Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40Yuexin Wang0Na Li1Guanda Liang2Luyang Wang3Xiaotian Zhang4Zhaohui Cui5Xiaoying Li6Sumei Zhang7Longxian Zhang8College of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityCollege of Veterinary Medicine, Henan Agricultural UniversityAbstract Background Cryptosporidium parvum is an apicomplexan zoonotic parasite causing the diarrheal illness cryptosporidiosis in humans and animals. To invade the host intestinal epithelial cells, parasitic proteins expressed on the surface of sporozoites interact with host cells to facilitate the formation of parasitophorous vacuole for the parasite to reside and develop. The gp40 of C. parvum, named Cpgp40 and located on the surface of sporozoites, was proven to participate in the process of host cell invasion. Methods We utilized the purified Cpgp40 as a bait to obtain host cell proteins interacting with Cpgp40 through the glutathione S-transferase (GST) pull-down method. In vitro analysis, through bimolecular fluorescence complementation assay (BiFC) and coimmunoprecipitation (Co-IP), confirmed the solid interaction between Cpgp40 and ENO1. In addition, by using protein mutation and parasite infection rate analysis, it was demonstrated that ENO1 plays an important role in the C. parvum invasion of HCT-8 cells. Results To illustrate the functional activity of Cpgp40 interacting with host cells, we identified the alpha-enolase protein (ENO1) from HCT-8 cells, which showed direct interaction with Cpgp40. The mRNA level of ENO1 gene was significantly decreased at 3 and 24 h after C. parvum infection. Antibodies and siRNA specific to ENO1 showed the ability to neutralize C. parvum infection in vitro, which indicated the participation of ENO1 during the parasite invasion of HCT-8 cells. In addition, we further demonstrated that ENO1 protein was involved in the regulation of cytoplasmic matrix of HCT-8 cells during C. parvum invasion. Functional study of the protein mutation illustrated that ENO1 was also required for the endogenous development of C. parvum. Conclusions In this study, we utilized the purified Cpgp40 as a bait to obtain host cell proteins ENO1 interacting with Cpgp40. Functional studies illustrated that the host cell protein ENO1 was involved in the regulation of tight junction and adherent junction proteins during C. parvum invasion and was required for endogenous development of C. parvum. Graphical Abstracthttps://doi.org/10.1186/s13071-024-06233-5Cryptosporidium parvumENO1Cpgp40GST pull-downInteraction |
| spellingShingle | Yuexin Wang Na Li Guanda Liang Luyang Wang Xiaotian Zhang Zhaohui Cui Xiaoying Li Sumei Zhang Longxian Zhang Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 Parasites & Vectors Cryptosporidium parvum ENO1 Cpgp40 GST pull-down Interaction |
| title | Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 |
| title_full | Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 |
| title_fullStr | Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 |
| title_full_unstemmed | Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 |
| title_short | Identification of host protein ENO1 (alpha-enolase) interacting with Cryptosporidium parvum sporozoite surface protein, Cpgp40 |
| title_sort | identification of host protein eno1 alpha enolase interacting with cryptosporidium parvum sporozoite surface protein cpgp40 |
| topic | Cryptosporidium parvum ENO1 Cpgp40 GST pull-down Interaction |
| url | https://doi.org/10.1186/s13071-024-06233-5 |
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