6533b834fe1ef96bd129d699
RESEARCH PRODUCT
Expression and purification of polyhistidine-tagged rotavirus NSP4 proteins in insect cells
Pilar López-andújarJavier BuesaJavier CuencaRebeca MontavaAna García-díazJesús Rodríguez-díazsubject
DiarrheaRotavirusGlycosylationInsectaImmunoprecipitationRecombinant Fusion ProteinsvirusesGenetic VectorsViral Nonstructural ProteinsProtein Engineeringmedicine.disease_causelaw.inventionMicelawRotavirusmedicineAnimalsHumansHistidinePolyacrylamide gel electrophoresisCells CulturedGlycoproteinsToxins Biologicalchemistry.chemical_classificationChemistryEndoplasmic reticulumbiochemical phenomena metabolism and nutritionMolecular biologyTransmembrane proteinBlotGene Expression RegulationRecombinant DNAElectrophoresis Polyacrylamide GelGlycoproteinBiotechnologydescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2003-10-11 | Protein Expression and Purification |