Structural properties of the ectodomain of hepatitis C virus E2 envelope protein

Abstract

We describe the structural and antigenic properties of a soluble form of hepatitis C virus E2 envelope protein ectodomain ending at residue 661 (E2661) which is obtained in large quantities in a baculovirus/insect cell system. The protein is secreted to the cellular medium by virus-infected cells. E2661 is glycosylated and possesses a high tendency to self-associate. In fact, analytical ultracentrifugation and size exclusion chromatography studies show that the purified protein is mainly composed of dimers, trimers and tetramers being the dimer the smallest species present in solution. The secondary structure was determined by deconvolution of the far-UV circular dichroism spectrum yielding 8% α-helix structure, 47% extended structure and 45% non-ordered structure. The near-UV CD spectrum is indicative of a folded structure. The fluorescence emission spectrum indicates that Trp residues occupy a relatively low hydrophobic environment. Finally, E2661 binds to a monoclonal conformation specific antibody and to antibodies present in human sera from HCV-positive patients. All these features suggest that the secreted protein possesses a native-like conformation. The use of this independent folding domain may contribute to shed light on the biology of HCV and could also be used as a vaccine in the prevention of HCV infection.