Epitope-distal Effects Accompany the Binding of Two Distinct Antibodies to Hepatitis B Virus Capsids

J.Z. Bereszczak, R.J. Rose, E. van Duijn, N.R. Watts, P.T. Wingfield, A.C. Steven, A.J.R. Heck

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Infection of humans by hepatitis B virus (HBV) induces the copious production of antibodies directed against the capsid protein (Cp). A large variety of anticapsid antibodies have been identified that differ in their epitopes. These data, and the status of the capsid as a major clinical antigen, motivate studies to achieve a more detailed understanding of their interactions. In this study, we focused on the Fab fragments of two monoclonal antibodies, E1 and 3120. E1 has been shown to bind to the side of outward-protruding spikes whereas 3120 binds to the “floor” region of the capsid, between spikes. We used hydrogen–deuterium exchange coupled to mass spectrometry (HDX-MS) to investigate the effects on HBV capsids of binding these antibodies. Conventionally, capsids loaded with saturating amounts of Fabs would be too massive to be readily amenable to HDX-MS. However, by focusing on the Cp protein, we were able to acquire deuterium uptake profiles covering the entire 149-residue sequence and reveal, in localized detail, changes in H/D exchange rates accompanying antibody binding. We find increased protection of the known E1 and 3120 epitopes on the capsid upon binding and show that regions distant from the epitopes are also affected. In particular, the α2a helix (residues 24–34) and the mobile C-terminus (residues 141–149) become substantially less solvent-exposed. Our data indicate that even at substoichiometric antibody binding an overall increase in the rigidity of the capsid is elicited, as well as a general dampening of its breathing motions.
Original languageEnglish
Pages (from-to)6504-6512
Number of pages9
JournalJournal of the American Chemical Society
Volume135
DOIs
Publication statusPublished - 2013

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