Fig. 6.
Effect of HR2 on 2B 60-mer insertion and topology. (A) Structural organization of full-length and truncated viroporin 2B constructs. (B) In vitro translation of truncated viroporin 2B 60-mer, 72-mer, 80-mer, and full-length (2B/CtGlyc) constructs in which a fused C-terminal N-glycosylation tag (rectangle) provides a simple readout for topology determination. The presence of RMs and nonglycosylated and glycosylated proteins (empty and black dots, respectively) is indicated. (C) In vitro glycosylation of truncated viroporin 2B-derived proteins. The level of glycosylation is quantified from SDS-PAGE gels by measuring the fraction of glycosylated (fg) versus glycosylated-plus-nonglycosylated (fng) molecules, using the equation p = fg/fg + fng. Data correspond to averages from at least four independent experiments, and error bars show standard deviations. (D) Topological models for 2B constructs. Nt, N terminus.