Characterization of modified hepatitis C virus E2 proteins expressed on the cell surface

The envelope proteins of hepatitis C virus (HCV) are the likely targets of neutralizing antibodies and their molecular and functional characterization is relevant for vaccine development. We previously showed that surface-exp ressed E2 is a better immunogen than intracellular E2 and, therefore, we we re interested in exploring more efficient ways to present E2 protein on the cell surface. We found that E2 targeted to the cell surface by replacement of its transmembrane domain did not bring El to the surface although El co uld be expressed independently on the cell surface if its transmembrane dom ain was similarly replaced. FAGS analysis suggested that E2 expressed on th e cell surface acquired its native conformation more efficiently when trunc ated at aa 661 than when truncated at aa 715. The shorter form of truncated E2 better retained the ability to bind the second extracellular loop (EC2) of CD81, the putative HCV receptor. Interestingly, deletion of the hyperva riable region 1 (HVR1) did not perceptibly alter E2 structure; cell-surface forms of E2 lacking the HVR1 remained reactive with conformation-sensitive MAbs and were able to bind recombinant EC2 of CD81, (C) 2000 Academic Pres s.