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Expression and structural properties of a chimeric protein based on the ectodomains of E1 and E2 hepatitis C virus envelope glycoproteins

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Tello, Daniel and Rodríguez-Rodríguez, Mar and Yélamos, Belén and Gómez-Gutiérrez, Julián and Ortega, Sara and Pacheco, Beatriz and Peterson, Darrell L. and Gavilanes, Francisco (2010) Expression and structural properties of a chimeric protein based on the ectodomains of E1 and E2 hepatitis C virus envelope glycoproteins. Protein Expression and Purification, 71 (2). pp. 123-131. ISSN 1046-5928; 1096-0279

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Official URL: http://www.sciencedirect.com/science/article/pii/S1046592810000513



Abstract

Hepatitis C virus encodes two enveloped glycoproteins, E1 and E2, which are involved in viral attachment and entry into target cells. We have obtained in insect cells infected by recombinant baculovirus a chimeric secreted recombinant protein, E1341E2661, containing the ectodomains of E1 and E2. The described procedure allows the purification of approximately 2 mg of protein from 1 L of culture media. Sedimentation velocity experiments and SDS-PAGE in the absence of reducing agents indicate that the protein has a high tendency to self-associate, the dimer being the main species observed. All the oligomeric forms observed maintain a conformation which is recognized by the conformation-dependent monoclonal antibody H53 directed against the E2 ectodomain. The spectroscopic properties of E1341E2661 are those of a three-dimensionally structured protein. Moreover, the chimeric protein is able to bind to human antibodies present in HCV-positive human sera. Accordingly, this chimeric soluble polypeptide chain may be a valuable tool to study the structure-function relationship of HCV envelope proteins.


Item Type:Article
Uncontrolled Keywords:Hepatitis C Virus, envelope protein, E1, E2, baculovirus, glycosylation
Subjects:Sciences > Chemistry > Biochemistry
ID Code:33661
Deposited On:19 Oct 2015 11:15
Last Modified:19 Oct 2015 11:15

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