FIG. 8.

NS2 plays a central role in coordinating interaction between the E1-E2 glycoprotein and NS3-4A enzyme complexes. (A) Structure of Jc1/E2(AP) and ΔNS2. Constructs are labeled as in Fig. 5A. (B) NS2 is required for E2-NS3 interaction. Protein lysates were prepared from Huh-7.5(BirA) cells transfected with the indicated constructs at 48 h postelectroporation. NS3 present in lysates (Total) or eluates from streptavidin beads (Bound) was separated by SDS-PAGE, transferred to PVDF membranes, and detected by immunoblotting. (C) NS2 mutations inhibit E2-NS3 interaction. Huh-7.5(BirA) cells were transfected with Jc1/E2(AP) or Jc1/E2(AP) containing the indicated NS2 mutations, and lysates were prepared 48 h postelectroporation. E2(AP)-associated NS3 protein (Bound) was detected as in Fig. 2B and 8A.