Universidad Complutense de Madrid
E-Prints Complutense

Production and Characterization of the Ectodomain of E2 Envelope Glycoprotein of Hepatitis C Virus Folded in the Presence of Full-length E1 glycoprotein

Impacto

Downloads

Downloads per month over past year



Ortega-Atienza, Sara and Lombana, Laura and Gómez-Gutiérrez, Julián and Yélamos, Belén and Peterson, Darrell L. and Gavilanes, Francisco (2014) Production and Characterization of the Ectodomain of E2 Envelope Glycoprotein of Hepatitis C Virus Folded in the Presence of Full-length E1 glycoprotein. Protein Expression and Purification, 104 . pp. 20-25. ISSN 1046-5928; 1096-0279

[img]
Preview
PDF
388kB

Official URL: http://www.sciencedirect.com/science/article/pii/S104659281400206X



Abstract

Hepatitis C virus (HCV) envelope glycoproteins, E1 and E2, are involved in the first steps of virus infection. The E2 ectodomain can be produced as an isolated form (E2661). However, there is some concern about its proper conformation and the role that E1 can play as a chaperone for the folding of E2. In order to verify this fact we have expressed a chimeric protein (E1tmbE2) based on the full-length E1 sequence followed by the E2 ectodomain using the baculovirus-insect cells system. The E2 ectodomain is folded in the presence of the E1, proteolytically processed by cellular proteases and secreted to cell culture media (E2661p), while the E1 protein is retained into the cell due to its transmembrane sequence. The purification of E2661p from culture media was facilitated by a His tag introduced in its amino terminus. Both E2661 and E2661p glycoproteins shared very similar structural features, monitored by spectroscopic and antigenic studies. Moreover, their functional properties, tested by means of CD81 binding, were almost indistinguishable, indicating that the E2 ectodomain constitutes an independent folding unit.


Item Type:Article
Uncontrolled Keywords:Hepatitis C virus envelope proteins; protein structure; E2 glycoprotein; E2 ectodomain
Subjects:Sciences > Chemistry > Biochemistry
ID Code:33668
Deposited On:19 Oct 2015 11:15
Last Modified:01 Jan 2016 00:01

Origin of downloads

Repository Staff Only: item control page