Complete set of binding and functional data for 545 HCV E1E2 mutations.
Complete set of binding and functional data for 545 HCV E1E2 mutations. To understand how HCV E1E2 folds and functions on an amino acid level, we generated an alanine scan library of mutants covering E1E2, using a parental expression construct for E1E2 (HCV genotype 1a, strain H77) with a V5 tag at the E2 C-terminus, and introduced an alanine substitution at each of 545 E1E2 residues. Existing alanine residues were changed to serine. All clones (545/555 residues, 98% coverage) were individually transfected intoHEK-293T cells and tested by flow cytometry for binding to 37 different MAbs and to a soluble fragment of CD81 (CD81-LEL). Data were obtained with each E1E2 mutant for the ability to support MAb binding, CD81 LEL binding, E1E2 expression, HCVpp budding, and HCVpp infectivity in HEK293T cells expressing Claudin 1 (CLDN1). Values are expressed as % of values obtained with WT E1E2 (defined as 100%). Some of the data have been shown in a previous examination of HCV E1E2 structure (Gopal et al., 2017 PLoS Pathog); the data set here extends this original data set to additional MAbs (including E1 MAbs) and tests the infectivity of each mutant. The line between A383 and E384 indicates the E1E2 junction.