Expression and purification of polyhistidine-tagged rotavirus NSP4 proteins in insect cells
| Autor: | Pilar López-Andújar, Javier Buesa, Javier Cuenca, Rebeca Montava, Ana García-Díaz, Jesús Rodríguez-Díaz |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Diarrhea
Rotavirus Glycosylation Insecta Immunoprecipitation Recombinant Fusion Proteins viruses Genetic Vectors Viral Nonstructural Proteins Protein Engineering medicine.disease_cause law.invention Mice law medicine Animals Humans Histidine Polyacrylamide gel electrophoresis Cells Cultured Glycoproteins Toxins Biological chemistry.chemical_classification Chemistry Endoplasmic reticulum biochemical phenomena metabolism and nutrition Molecular biology Transmembrane protein Blot Gene Expression Regulation Recombinant DNA Electrophoresis Polyacrylamide Gel Glycoprotein Biotechnology |
| Zdroj: | Protein Expression and Purification. 31:207-212 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/s1046-5928(03)00166-9 |
| Popis: | The rotavirus nonstructural NSP4 protein, a transmembrane endoplasmic reticulum-specific glycoprotein, has been described as the first viral enterotoxin. Purified NSP4 or a peptide corresponding to NSP4 residues 114-135 induces diarrhea in young mice. NSP4 has a membrane-destabilizing activity and causes an increase in intracellular calcium levels and chloride secretion by a calcium-dependent signalling pathway in eucaryotic cells. In this study, four recombinant baculoviruses were generated expressing the rotavirus NSP4 glycoprotein from the human strains Wa and Ito, the porcine strain OSU, and the simian strain SA11, which belong to two different NSP4 genotypes, A and B. The recombinant glycoproteins, expressed as polyhistidine-tagged molecules, were analyzed by Western blotting and immunoprecipitation. Newborn mice responded with diarrhea after inoculation with each of the recombinant NSP4 proteins. |
| Databáze: | OpenAIRE |
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