Physical and immunological characterization of a recombinant secreted form of the membrane protein encoded by the vaccinia virus L1R gene.
We reported that immunization with recombinant proteins derived from vaccinia virus (VV) particles could provide protection against infection. Here we describe the physical and antigenic properties of the L1R membrane protein. The recombinant protein (L1R(185t)) was secreted as a monomer and correct folding was suggested by the presence of three intramolecular disulfide bonds and binding to conformation-specific monoclonal antibodies (MAbs). Furthermore, anti-L1R(185t) rabbit antisera exhibited potent virus-neutralizing activity against the IMV form of VV. We raised six MAbs against L1R(185t). Three recognized linear epitopes (residues 118--128) and neutralized IMV infectivity. These MAbs blocked binding of each other to L1R(185t) but failed to block binding of two previously described neutralizing anti-L1R MAbs, 7D11 and 2D5. The latter two antibodies blocked each other in binding L1R(185t). Thus, two antigenic sites on L1R overlap functional domains and based on recent structural studies these are found in accessible regions of the IMV L1R protein.
Aldaz-Carroll
Lydia
L
Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104-6002, USA. aldaz@biochem.dental.upenn.edu
Whitbeck
J Charles
JC
Ponce de Leon
Manuel
M
Lou
Huan
H
Pannell
Lewis K
LK
Lebowitz
Jacob
J
Fogg
Christiana
C
White
Christine L
CL
Moss
Bernard
B
Cohen
Gary H
GH
Eisenberg
Roselyn J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D005814:Genes, Viral; D006031:Glycosylation; D066298:In Vitro Techniques; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D000939:Epitopes; C087715:L1R protein, Vaccinia virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins