title: Cross-linking of rotavirus outer capsid protein VP7 by antibodies or disulfides inhibits viral entry.
abstract: Antibodies that neutralize rotavirus infection target outer coat proteins VP4 and VP7 and inhibit viral entry. The structure of a VP7-Fab complex (S. T. Aoki, et al., Science 324:1444-1447, 2009) led us to reclassify epitopes into two binding regions at inter- and intrasubunit boundaries of the calcium-dependent trimer. It further led us to show that antibodies binding at the intersubunit boundary inhibit uncoating of the virion outer layer. We have now tested representative antibodies for each of the defined structural epitope regions and find that antibodies recognizing epitopes in either binding region neutralize by cross-linking VP7 trimers. Antibodies that bind at the intersubunit junction neutralize as monovalent Fabs, while those that bind at the intrasubunit region require divalency. The VP7 structure has also allowed us to design a disulfide cross-linked VP7 mutant which recoats double-layered particles (DLPs) as efficiently as does wild-type VP7 but which yields particles defective in cell entry as determined both by lack of infectivity and by loss of α-sarcin toxicity in the presence of recoated particles. We conclude that dissociation of the VP7 trimer is an essential step in viral penetration into cells.
authors.affiliation: Laboratory of Molecular Medicine, Children's Hospital, 3 Blackfan Circle, Boston, MA 02115, USA.
authors.forename: Scott T
authors.initials: ST
authors.lastname: Aoki
authors.forename: Shane D
authors.initials: SD
authors.lastname: Trask
authors.forename: Barbara S
authors.initials: BS
authors.lastname: Coulson
authors.forename: Harry B
authors.initials: HB
authors.lastname: Greenberg
authors.forename: Philip R
authors.initials: PR
authors.lastname: Dormitzer
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Harrison
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D036022:Capsid Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D004220:Disulfides / Q000378:metabolism; D008253:Macaca mulatta; D012401:Rotavirus / Q000502:physiology*; D053586:Virus Internalization
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D004220:Disulfides; C050779:VP7 protein, Rotavirus