Protein disulfide isomerase and heat shock cognate protein 70 interactions with rotavirus structural proteins using their purified recombinant versions
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| Format: | Artículo científico |
| Language: | en |
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Universidad Nacional de Colombia
2016
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| _version_ | 1876444082080841728 |
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| author | Luz Y. Moreno |
| author_facet | Luz Y. Moreno |
| contents | Protein disulfide isomerase and heat shock cognate protein 70 interactions with rotavirus structural proteins using their purified recombinant versions Luz Y. Moreno Carlos A. Guerrero Orlando Acosta Biología Rotavirus recombinant proteins protein disulfide isomerase heat shock cognate protein 70 Introduction. Rotavirus entry into cells seems to be mediated by sequential interactions between viral structural proteins and some cell surface molecules. However, the mechanisms by which rotavirus infects target cell are still not well understood. There is some evidence showing that rotavirus structural proteins VP5* and VP8* interact with some cell surface molecules. The availability of recombinant rotavirus structural proteins in sufficient quantity has become very important for the identification of the specific virus-cell receptor interactions during the early events of the infectious process. Objective. The aim of the present work is to perform an analysis of the interactions between recombinant rotavirus structural proteins VP5*, VP8* and VP6, and cellular proteins Hsc70 and PDI using their purified recombinant versions. Materials and methods. Rotavirus recombinant VP5* and VP8*, and cellular recombinant proteins Hsc70 and PDI were expressed in E. coli BL21(DE3) while VP6 was expressed in recombinant vaccinia virus-transfected MA104 cells. The interaction between rotavirus and cellular proteins was studied using ELISA, co-immunoprecipitation and SDS-PAGE/Western blotting analysis. Results. The optimal conditions for expression of recombinant proteins were determined and antibodies were raised against them. The findings suggested that viral proteins rVP5* and rVP6 interact with Hsc70 and PDI in vitro. These viral recombinant proteins were also found to interact with raft-associated Hsc70 in a cell culture system. The treatment of cells with either rVP6 or DLPs produced significantly inhibition of rotavirus infection. Conclusion. The results allow us to conclude that rVP5* and rVP6 interact with Hsc70 and PDI during the rotavirus infection process. 2016 artículo científico 0123-3475 https://www.redalyc.org/articulo.oa?id=77645907004 https://www.redalyc.org/journal/776/77645907004/ https://www.redalyc.org/journal/776/77645907004/html/ https://www.redalyc.org/journal/776/77645907004/77645907004.epub https://www.redalyc.org/journal/776/77645907004/movil en http://www.redalyc.org/revista.oa?id=776 Revista Colombiana de Biotecnología application/pdf Universidad Nacional de Colombia Revista Colombiana de Biotecnología (Colombia) Num.1 Vol.XVIII |
| format | Artículo científico |
| id | redalyc_77645907004 |
| institution | Redalyc |
| language | en |
| publishDate | 2016 |
| publisher | Universidad Nacional de Colombia |
| spellingShingle | Protein disulfide isomerase and heat shock cognate protein 70 interactions with rotavirus structural proteins using their purified recombinant versions Luz Y. Moreno Biología Rotavirus recombinant proteins protein disulfide isomerase heat shock cognate protein 70 Protein disulfide isomerase and heat shock cognate protein 70 interactions with rotavirus structural proteins using their purified recombinant versions Luz Y. Moreno Carlos A. Guerrero Orlando Acosta Biología Rotavirus recombinant proteins protein disulfide isomerase heat shock cognate protein 70 Introduction. Rotavirus entry into cells seems to be mediated by sequential interactions between viral structural proteins and some cell surface molecules. However, the mechanisms by which rotavirus infects target cell are still not well understood. There is some evidence showing that rotavirus structural proteins VP5* and VP8* interact with some cell surface molecules. The availability of recombinant rotavirus structural proteins in sufficient quantity has become very important for the identification of the specific virus-cell receptor interactions during the early events of the infectious process. Objective. The aim of the present work is to perform an analysis of the interactions between recombinant rotavirus structural proteins VP5*, VP8* and VP6, and cellular proteins Hsc70 and PDI using their purified recombinant versions. Materials and methods. Rotavirus recombinant VP5* and VP8*, and cellular recombinant proteins Hsc70 and PDI were expressed in E. coli BL21(DE3) while VP6 was expressed in recombinant vaccinia virus-transfected MA104 cells. The interaction between rotavirus and cellular proteins was studied using ELISA, co-immunoprecipitation and SDS-PAGE/Western blotting analysis. Results. The optimal conditions for expression of recombinant proteins were determined and antibodies were raised against them. The findings suggested that viral proteins rVP5* and rVP6 interact with Hsc70 and PDI in vitro. These viral recombinant proteins were also found to interact with raft-associated Hsc70 in a cell culture system. The treatment of cells with either rVP6 or DLPs produced significantly inhibition of rotavirus infection. Conclusion. The results allow us to conclude that rVP5* and rVP6 interact with Hsc70 and PDI during the rotavirus infection process. 2016 artículo científico 0123-3475 https://www.redalyc.org/articulo.oa?id=77645907004 https://www.redalyc.org/journal/776/77645907004/ https://www.redalyc.org/journal/776/77645907004/html/ https://www.redalyc.org/journal/776/77645907004/77645907004.epub https://www.redalyc.org/journal/776/77645907004/movil en http://www.redalyc.org/revista.oa?id=776 Revista Colombiana de Biotecnología application/pdf Universidad Nacional de Colombia Revista Colombiana de Biotecnología (Colombia) Num.1 Vol.XVIII |
| title | Protein disulfide isomerase and heat shock cognate protein 70 interactions with rotavirus structural proteins using their purified recombinant versions |
| topic | Biología Rotavirus recombinant proteins protein disulfide isomerase heat shock cognate protein 70 |
| url | https://www.redalyc.org/articulo.oa?id=77645907004 https://www.redalyc.org/journal/776/77645907004/ https://www.redalyc.org/journal/776/77645907004/html/ https://www.redalyc.org/journal/776/77645907004/77645907004.epub https://www.redalyc.org/journal/776/77645907004/movil |