Sindbis virus conformational changes induced by a neutralizing anti-E1 monoclonal antibody.
A rare Sindbis virus anti-E1 neutralizing monoclonal antibody, Sin-33, was investigated to determine the mechanism of in vitro neutralization. A cryoelectron microscopic reconstruction of Sindbis virus (SVHR) neutralized with FAb from Sin-33 (FAb-33) revealed conformational changes on the surface of the virion at a resolution of 24 A. FAb-33 was found to bind E1 in less than 1:1 molar ratios, as shown by the absence of FAb density in the reconstruction and stoichiometric measurements using radiolabeled FAb-33, which determined that about 60 molecules of FAb-33 bound to the 240 possible sites in a single virus particle. FAb-33-neutralized virus particles became sensitive to digestion by endoproteinase Glu-C, providing further evidence of antibody-induced structural changes within the virus particle. The treatment of FAb-33-neutralized or Sin-33-neutralized SVHR with low pH did not induce the conformational rearrangements required for virus membrane-cell membrane fusion. Exposure to low pH, however, increased the amount of Sin-33 or FAb-33 that bound to the virus particles, indicating the exposure of additional epitopes. The neutralization of SVHR infection by FAb-33 or Sin-33 did not prevent the association of virus with host cells. These data are in agreement with the results of previous studies that demonstrated that specific antibodies can inactivate the infectious state of a metastable virus in vitro by the induction of conformational changes to produce an inactive structure. A model is proposed which postulates that the induction of conformational changes in the infectious state of a metastable enveloped virus may be a general mechanism of antibody inactivation of virus infectivity.
Hernandez
Raquel
R
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27608, USA. raquel_hernandez@ncsu.edu
Paredes
Angel
A
Brown
Dennis T
DT
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D006224:Cricetinae; D000939:Epitopes; D006863:Hydrogen-Ion Concentration; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007668:Kidney / Q000166:cytology; D008562:Membrane Glycoproteins / Q000276:immunology*; D009500:Neutralization Tests; D010449:Peptide Mapping; D011485:Protein Binding; D011487:Protein Conformation; D012845:Sindbis Virus / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D014759:Viral Envelope Proteins / Q000276:immunology*; D014771:Virion / Q000502:physiology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; C022012:glycoprotein E1, Sindbis virus