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An optimized hepatitis C virus E2 glycoprotein core adopts a functional homodimer that efficiently blocks virus entry.

McCaffrey K, Boo I, Owczarek CM, Hardy MP, Perugini MA, Fabri L, Scotney P, Poumbourios P, Drummer HE

Abstract

The hepatitis C virus (HCV) envelope glycoprotein E2 is the major target of broadly neutralizing antibodies in vivo and is the focus of efforts in the rational design of a universal B cell vaccine against HCV. The E2 glycoprotein exhibits a high degree of amino-acid variability which localizes to three discrete regions: hypervariable region 1 (HVR1), hypervariable region 2 (HVR2) and the intergenotypic variable region (igVR). All three variable regions contribute to immune evasion and/or isolate-specific structural variations, both important considerations for vaccine design. High resolution structural definition of the intact HCV envelope glycoprotein complex containing E1 and E2 remains to be elucidated whilst crystallographic structures of a recombinant E2 ectodomain failed to resolve HVR1, HVR2 and a major neutralization determinant adjacent to HVR1. To obtain further information on E2, we characterized the role of all three variable regions in E2 ectodomain folding and function in the context of a recombinant ectodomain fragment (rE2). We report that removal of the variable regions accelerates binding to the major host-cell receptor CD81 and that simultaneous deletion of HVR2 and the igVR is required to maintain wild-type CD81-binding characteristics. The removal of the variable regions also rescued the ability of rE2 to form a functional homodimer. We propose that the phenotype of the D123 dimer provides novel insights into the role of the variable motifs in the higher order assembly of the E2 ectodomain and may have implications for E1E2 structure on the virion surface. IMPORTANCE: Hepatitis C virus (HCV) infection affects ∼2% of the population globally and no vaccine is available. HCV is a highly variable virus and understanding the presentation of key antigenic sites at the virion surface is important for the design of a universal vaccine. This study investigates the role of three surface-exposed variable regions in E2 glycoprotein folding and function in the context of a recombinant soluble ectodomain. Our data demonstrates the variable motifs modulate binding of the E2 ectodomain to the major host-cell receptor CD81 and impact on the formation of an E2 homodimer with high affinity binding to CD81.

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(We gratefully acknowledge funding support from the National Health and Medical Research Council through project grants 543113, 1020175 and 1080045. HED was the recipient of RD Wright Fellowship 433929 and is currently supported from a Senior Research Fellowship 1041897 from the NHMRC. KM was the recipient of Dora Lush Postgraduate Biomedical Scholarship 433913 and is currently supported by a Marie Curie International Postdoctoral Fellowship from the European Commission. The authors gratefully acknowledge the contribution to this work of the Victorian Operational Infrastructure Support Program received by the Burnet Institute)

Publication

  • Journal: Journal of Virology
  • Published: 14/02/2017
  • Volume: 91
  • Issue: 5
  • Pagination: e01668

Authors

Program

Health Issue

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