Epitope mapping on fragments of beet necrotic yellow vein virus coat protein

• Published in: Journal of general virology 1992-03, Vol.73 (3), p.695-700
• Main Authors: Commandeur, U, Koenig, R, Lesemann, D.-E, Torrance, L, Burgermeister, W, Liu, Y, Schots, A, Alric, M, Grassi, G
• Format: Article
• Language: English
• Subjects: Antigenic determinants, haptens, artificial antigens; Antigens; beet necrotic yellow vein virus; Biological and medical sciences; Blotting, Western; Capsid - immunology; Epitopes - immunology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Gene Expression; Models, Biological; Molecular immunology; Peptide Fragments - immunology; Plant Viruses - immunology; Plants, Edible - microbiology; Recombinant Proteins - immunology
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-73-3-695

1 Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Biochemie und Pflanzenvirologie, Messeweg 11, D-3300 Braunschweig, Germany 2 Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, U.K. 3 Beijing Agricultural University, Beijing, China 4 Laboratory for Monoclonal Antibodies, NL-6700 GW Wageningen, The Netherlands 5 Laboratoire de Biologie Cellulaire et Moléculaire, F-63170 Aubiere, France and 6 Istituto Sperimentale per le Colture Industriale, S.O.P. de Rovigo, I-45000 Rovigo, Italy The location of five SDS-stable epitopes on the coat protein (CP) of beet necrotic yellow vein virus was determined by reacting Escherichia coli -expressed free CP, as well as fusion proteins (FP) containing fragments of the CP, with polyclonal and monoclonal antibodies on Western blots. Epitope 1, which has previously been found to be exposed on only one extremity of the virus particle, was located in the region between amino acids (aa) 1 and 7, i.e. on the N terminus of the CP. It was blocked when the N terminus of the CP was linked to a portion of the -galactosidase sequence in an FP. Epitope 3, which has previously been found to be exposed on the opposite extremity of the particle, was located in the region between aa 37 and 59. Epitope 4, which is exposed along the entire length of the particle, occurs on the C terminus of CP (aa 183 to 188). Two previously unknown epitopes were identified in the regions between aa 115 and 125 and 125 and 140, respectively. The former was located on the same extremity of the particle as epitope 3, the latter became accessible only after denaturation of the particle. Nothing is known about the probably non-adjacent aa sequences that participate in the formation of the two SDS-labile epitopes (epitopes 2 and 5) which are found on one extremity and along the entire length of the particle, respectively. Received 11 September 1991; accepted 19 November 1991.