Biopolym. Cell. 2000; 16(4):312-319.
http://dx.doi.org/10.7124/bc.000577
Viruses and Cell
Analysis of antigenic structure of Potato Virus M Ukrainian strains
1Viter S. S., 1Tkachenko T. Yu., 2Kolomietz L. P., 1Radavsky Yu. L.
  1. Institute of Bioorganic Chemistry and Petrochemistry, NAS of Ukraine
    1, Murmans'ka Str., Kyiv, Ukraine, 02094
  2. Institute of Agricultural Microbiology and agricultural production NAAS
    97, Shevchenko Str., Chernihiv, Ukraine, 14027

Abstract

The antigenic structure of two Ukrainian strains V1 and U7 of Potato Virus M (PVM) was studied by using three mouse monoclonal antibodies (MAbs). Tryptic fragments of PVM coat protein (CP) 22Glu-Lys35 and 22Gly-Lys35 were found to be recognised by two MAbs, M6D5 and M9G1. The amino acid substitution Glu → Gly at the position 22 of PVM 117 coat protein did not affect antibody binding to both tryptic fragments and PVM CP. Mob M9GJ interfered with MAb M4C1 in dot immunobinding assay. Synthetic peptide P14, corresponding to tryptic fragment 22Glu-Lys35, inhibited the interaction of MAb M6D5 with CP and peptide P14 more effectively than that of MAb M9G1. The modification of the side chain positive charge of Lys35 to negative one in tryptic peptides 22Glu-Lys35 and 22Gly-Lys35 using citraconic anhydride resulted in a two-fold increase of the reaction of MAb M9G1 and slightly reduce the interaction of MAb M6D5 with both fragments. On the basis of these results, it was concluded that three PVM-specific epitopes are located at the N-terminal region and form original immunogenic site of PVM coat protein. MAbs M6D5 and M9G1 recognise sequentially overlapping epitopes and the common site for both epitopes is presented in the fragment 22Glu/Gly-Lys35. MAbs M4C1 and M9G1 recognise either overlapping, or conformational approximated epitopes.
Full text: (PDF, in English)

References

[1] Tavantzis SM. Improved purification of two potato carlaviruses. Phytopathology. 1983; 73(2): 190-4.
[2] Tavantzis SM. Physicochemical properties of potato virus M. Virology. 1984;133(2):427-30.
[3] Zavriev SK, Kanyuka KV, Levay KE. The genome organization of potato virus M RNA. J Gen Virol. 1991;72 ( Pt 1):9-14.
[4] Gramstat A, Courtpozanis A, Rohde W. The 12 kDa protein of potato virus M displays properties of a nucleic acid-binding regulatory protein. FEBS Lett. 1990;276(1-2):34-8.
[5] Foster GD. The structure and expression of the genome of carlaviruses. Res Virol. 1992;143(2):103-12.
[6] Wetter C, Milne RG. Carlaviruses. Handbook of plant virus infections and comparative diagnosis. Ed. E. Kurstak. Amsterdam; New York; Oxford: Elsevier. North-Holland Bio­chemical press, 1981: 685-730.
[7] Viter SS, Raudsepp RA, Gavrish OG, Radavsky YuL. Localization of epitope on potato virus M coat protein. Doklady Akad Nauk Ukrainy. 1992;(9):162-4.
[8] Foster GD, Mills PR. Analysis of the coat proteins of the ordinary and Andean strains of potato virus S and antigenic comparisons of carlaviruses. Acta Virol. 1992;36(2):184-90.
[9] Cerovsk? N, Filigarov? M, Subr Z. Partial antigenic characterization of potato virus S (Andean strain) by monoclonal antibodies. Acta Virol. 1996;40(1):23-6.
[10] Adams AN, Barbara DJ. The use of F(ab‘) 2 -based ELISA to detect serological relationships among carlaviruses . Ann Appl Biol. 1982;101(3):495–500.
[11] J?rvek?lg L, S?ber J, Sinij?rv R, Toots I, Saarma M. Time-resolved fluoroimmunoassay of potato virus M with monoclonal antibodies. Ann Appl Biol. 1989;114(2):279–91.
[12] Nikolaeva OV, Novikov VK Atabekov IG, Kaftanova AS. Changes antigenic properties of potato virus M at the progressive degradation of structural protein in the process of separation and storage. S-H. biology. 1985;10.:75-8.
[13] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[14] Wu GJ, Bruening G. Two proteins from cowpea mosaic virus. Virology. 1971;46(3):596-612.
[15] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
[16] Barany G, Merrifield RB. Solid-phase peptide synthesis. Peptides. Eds E. Gross, J. Meienhoter. New York: Acad, press, 1980. Vol. 2: 1-284.
[17] Pitt-Rivers R, Impiombato FS. The binding of sodium dodecyl sulphate to various proteins. Biochem J. 1968;109(5):825-30.
[18] Tozzini AC, Ek B, Palva ET, Hopp HE. Potato virus X coat protein: a glycoprotein. Virology. 1994;202(2):651-8.
[19] Muller S, Plaue S, Couppez M, Van Regenmortel MH. Comparison of different methods for localizing antigenic regions in histone H2A. Mol Immunol. 1986;23(6):593-601.
[20] Joisson C, Kuster F, Plau? S, Van Regenmortel MH. Antigenic analysis of bean pod mottle virus using linear and cyclized synthetic peptides. Arch Virol. 1993;128(3-4):299-317.
[21] Rajashankar KR, Ramakumar S. Pi-turns in proteins and peptides: Classification, conformation, occurrence, hydration and sequence. Protein Sci. 1996;5(5):932-46.
[22] Shukla DD, Strike PM, Tracy SL, Gough KH, Ward CW. The N and C Termini of the Coat Proteins of Potyviruses Are Surface-located and the N Terminus Contains the Major Virus-specific Epitopes. J Gen Virol. 1988;69(7):1497–508.
[23] Radavsky YuL, Viter SS, Turova IP, Zaitseva LS, J?rvek?lg LV, Saarma MJu, Grebenshchikov NI, Baratova LA. Antigenic structure of the potato virus X coat protein. II. Localization of antigenic determinant(s) at the N-terminus of the protein. Bioorg Khim. 1989; 15 (5):615-9
[24] Baratova LA, Grebenshchikov NI, Dobrov EN, Gedrovich AV, Kashirin IA, Shishkov AV, Efimov AV, J?rvek?lg L, Radavsky YL, Saarma M. The organization of potato virus X coat proteins in virus particles studied by tritium planigraphy and model building. Virology. 1992;188(1):175-80.