Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum
Abstract Background Cryptosporidium spp. are important diarrhea-causing pathogens in humans and animals. Comparative genomic analysis indicated that Cryptosporidium-specific MEDLE family proteins may contribute to host adaptation of Cryptosporidium spp., and a recent study of one member of this fami...
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| Format: | Article |
| Language: | English |
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BMC
2018-05-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-018-2889-2 |
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| author | Jilan Fei Haizhen Wu Jiayuan Su Chanchan Jin Na Li Yaqiong Guo Yaoyu Feng Lihua Xiao |
| author_facet | Jilan Fei Haizhen Wu Jiayuan Su Chanchan Jin Na Li Yaqiong Guo Yaoyu Feng Lihua Xiao |
| author_sort | Jilan Fei |
| collection | DOAJ |
| description | Abstract Background Cryptosporidium spp. are important diarrhea-causing pathogens in humans and animals. Comparative genomic analysis indicated that Cryptosporidium-specific MEDLE family proteins may contribute to host adaptation of Cryptosporidium spp., and a recent study of one member of this family, CpMEDLE-2 encoded by cgd5_4590, has provided evidence supporting this hypothesis. In this study, another member of the protein family, CpMEDLE-1 of Cryptosporidium parvum encoded by cgd5_4580, which is distinct from CpMEDLE-2 and has no signature motif MEDLE, was cloned, expressed and characterized to understand its function. Methods CpMEDLE-1 was expressed in Escherichia coli and polyclonal antibodies against the recombinant CpMEDLE-1 protein were prepared in rabbits. Quantitative PCR was used to analyze the expression profile of cgd5_4580 in C. parvum culture. Immunofluorescence staining was used to locate CpMEDLE-1 expression in life-cycle stages, and in vitro neutralization assay with antibodies was adopted to assess the role of the protein in C. parvum invasion. Results The results indicated that cgd5_4580 had a peak expression at 2 h of C. parvum culture. CpMEDLE-1 was located in the mid-anterior region of sporozoites, probably within the dense granules. The neutralization efficiency of anti-CpMEDLE-1 antibodies was approximately 40%. Conclusions The differences in protein and gene expression profiles between CpMEDLE-1 and CpMEDLE-2 suggest that MEDLE proteins have different subcellular locations, are developmentally regulated, could be potentially involved in the transcriptional regulation of the expression of parasite or host proteins and may exert their functions in different stages of the invasion and development process. |
| first_indexed | 2024-12-10T22:27:55Z |
| format | Article |
| id | doaj.art-2ab1159770de4dc7bfb99924a9124b54 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-10T22:27:55Z |
| publishDate | 2018-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-2ab1159770de4dc7bfb99924a9124b542022-12-22T01:31:08ZengBMCParasites & Vectors1756-33052018-05-011111910.1186/s13071-018-2889-2Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvumJilan Fei0Haizhen Wu1Jiayuan Su2Chanchan Jin3Na Li4Yaqiong Guo5Yaoyu Feng6Lihua Xiao7State Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and TechnologySchool of Biotechnology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and TechnologyKey Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural UniversityKey Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural UniversityState Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and TechnologyKey Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural UniversityAbstract Background Cryptosporidium spp. are important diarrhea-causing pathogens in humans and animals. Comparative genomic analysis indicated that Cryptosporidium-specific MEDLE family proteins may contribute to host adaptation of Cryptosporidium spp., and a recent study of one member of this family, CpMEDLE-2 encoded by cgd5_4590, has provided evidence supporting this hypothesis. In this study, another member of the protein family, CpMEDLE-1 of Cryptosporidium parvum encoded by cgd5_4580, which is distinct from CpMEDLE-2 and has no signature motif MEDLE, was cloned, expressed and characterized to understand its function. Methods CpMEDLE-1 was expressed in Escherichia coli and polyclonal antibodies against the recombinant CpMEDLE-1 protein were prepared in rabbits. Quantitative PCR was used to analyze the expression profile of cgd5_4580 in C. parvum culture. Immunofluorescence staining was used to locate CpMEDLE-1 expression in life-cycle stages, and in vitro neutralization assay with antibodies was adopted to assess the role of the protein in C. parvum invasion. Results The results indicated that cgd5_4580 had a peak expression at 2 h of C. parvum culture. CpMEDLE-1 was located in the mid-anterior region of sporozoites, probably within the dense granules. The neutralization efficiency of anti-CpMEDLE-1 antibodies was approximately 40%. Conclusions The differences in protein and gene expression profiles between CpMEDLE-1 and CpMEDLE-2 suggest that MEDLE proteins have different subcellular locations, are developmentally regulated, could be potentially involved in the transcriptional regulation of the expression of parasite or host proteins and may exert their functions in different stages of the invasion and development process.http://link.springer.com/article/10.1186/s13071-018-2889-2Cryptosporidium parvumMEDLE familyInvasionNeutralizationExpression |
| spellingShingle | Jilan Fei Haizhen Wu Jiayuan Su Chanchan Jin Na Li Yaqiong Guo Yaoyu Feng Lihua Xiao Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum Parasites & Vectors Cryptosporidium parvum MEDLE family Invasion Neutralization Expression |
| title | Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum |
| title_full | Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum |
| title_fullStr | Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum |
| title_full_unstemmed | Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum |
| title_short | Characterization of MEDLE-1, a protein in early development of Cryptosporidium parvum |
| title_sort | characterization of medle 1 a protein in early development of cryptosporidium parvum |
| topic | Cryptosporidium parvum MEDLE family Invasion Neutralization Expression |
| url | http://link.springer.com/article/10.1186/s13071-018-2889-2 |
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