title: Antigenic analysis of the coat protein of beet necrotic yellow vein virus by means of monoclonal antibodies.
abstract: By means of monoclonal antibodies (MAbs), five (groups of) epitopes were identified on particles of beet necrotic yellow vein virus (BNYVV). Epitopes 1 and 2, which were located on the opposite extremities of virus particles, are discontinuous (SDS-labile) epitopes which were destroyed when the particles were treated with trypsin. Epitope 3 is a continuous (SDS-stable) epitope located at the same extremity as epitope 2. It was not destroyed when the particles were treated with trypsin and was present on an Escherichia coli-expressed fusion protein containing amino acids (aa) 1 to 103 of the BNYVV coat protein. The continuous epitope 4, which was located along the entire length of the particles, was found to be present on a fusion protein containing aa 104 to 188 of the BNYVV coat protein but not on trypsin-treated virus particles. In Western blots, these treated particles yielded two slightly smaller coat proteins which failed to react with MAbs specific for epitope 4 but did react with polyclonal antisera and MAbs specific for epitope 3. BNYVV coat protein has a trypsin cleavage site on the carboxyl side of arginine in position 182, so it is therefore suggested that epitope 4 is located on the exposed C terminus, which is composed of aa 183 to 188. Epitope 5 was also located along the entire length of the particles but in a more uneven distribution than epitope 4. This may be because it is a discontinuous epitope that is very sensitive to subtle changes in protein conformation.
authors.affiliation: Biologische Bundesanstalt für Land- und Forstwirtschaft, Institute für Viruskrankheiten der Pflanzen und Biochemie, Braunschweig, F.R.G.
authors.forename: R
authors.initials: R
authors.lastname: Koenig
authors.forename: U
authors.initials: U
authors.lastname: Commandeur
authors.forename: D E
authors.initials: DE
authors.lastname: Lesemann
authors.forename: W
authors.initials: W
authors.lastname: Burgermeister
authors.forename: L
authors.initials: L
authors.lastname: Torrance
authors.forename: G
authors.initials: G
authors.lastname: Grassi
authors.forename: M
authors.initials: M
authors.lastname: Alric
authors.forename: J
authors.initials: J
authors.lastname: Kallerhoff
authors.forename: A
authors.initials: A
authors.lastname: Schots
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D015153:Blotting, Western; D002213:Capsid / Q000276:immunology*; D008854:Microscopy, Electron; D010942:Plant Viruses / Q000145:classification / Q000276:immunology* / Q000648:ultrastructure; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014357:Trypsin / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D014357:Trypsin