We separated E1 and E2 of hepatitis C virus (HCV) genotypes 1a, 1b, and 2a into two individual expression plasmids and replaced the transmembrane domains of 1b and 2a E1 and E2 with that of genotype 1a. The complementation features of El and E2 as well as the contributions of both the ecto- and transmembrane domains to the formation of the E1E2 complex were evaluated using the HCV peudoparticle(s) (HCVpp(s)) system. We demonstrated that 1aE2 could not only complement its native 1aE1, but could also complement 1bE1 as well: in genotype 1b, glycoprotein complex formation is primarily dependent on the overall biological characteristics of the intact native E1 and E2; in genotype 2a, although the interaction of intact native E1 and E2 is critical for the formation of the glycoprotein complex, the ectodomain made a greater contribution than that of the transmembrane domain. Our study provides valuable findings regarding HCV E1 and E2 biology and will be of use in both anti-HCV strategy and understanding oil the mechanisms of coinfection of different HCV strains. (C) 2009 Elsevier Inc. All rights reserved.