title: Analysis of the epitope structure of Plum pox virus coat protein.
abstract: Typing of the particular Plum pox virus (PPV) strain responsible in an outbreak has important practical implications and is frequently performed using strain-specific monoclonal antibodies (MAbs). Analysis in Western blots of the reactivity of 24 MAbs to a 112-amino-acid N-terminal fragment of the PPV coat protein (CP) expressed in Escherichia coli showed that 21 of the 24 MAbs recognized linear or denaturation-insensitive epitopes. A series of eight C-truncated CP fragments allowed the mapping of the epitopes recognized by the MAbs. In all, 14 of them reacted to the N-terminal hypervariable region, defining a minimum of six epitopes, while 7 reacted to the beginning of the core region, defining a minimum of three epitopes. Sequence comparisons allowed the more precise positioning of regions recognized by several MAbs, including those recognized by the 5B-IVIA universal MAb (amino acids 94 to 100) and by the 4DG5 and 4DG11 D serogroup-specific MAbs (amino acids 43 to 64). A similar approach coupled with infectious cDNA clone mutagenesis showed that a V74T mutation in the N-terminus of the CP abolished the binding of the M serogroup-specific AL MAb. Taken together, these results provide a detailed positioning of the epitopes recognized by the most widely used PPV detection and typing MAbs.
authors.affiliation: Universite de Bordeaux, Villenave d'Ornon Cedex, France. tc@bordeaux.inra.fr
authors.forename: Thierry
authors.initials: T
authors.lastname: Candresse
authors.forename: Pilar
authors.initials: P
authors.lastname: Saenz
authors.forename: Juan Antonio
authors.initials: JA
authors.lastname: García
authors.forename: Donato
authors.initials: D
authors.lastname: Boscia
authors.forename: Milan
authors.initials: M
authors.lastname: Navratil
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Gorris
authors.forename: Mariano
authors.initials: M
authors.lastname: Cambra
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000821:virology*; D017801:Plum Pox Virus / Q000235:genetics / Q000276:immunology*; D027861:Prunus / Q000821:virology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D011994:Recombinant Proteins; C095320:coat protein, Plum pox virus