Biopolym. Cell. 2001; 17(5):417-422.
http://dx.doi.org/10.7124/bc.0005CE
Structure and Function of Biopolymers
Characterization of Potato Virus M epitopes with the use of synthetic peptides
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

As a result of thermolysin hydrolysis of a coat protein (CP) of Potato Virus M Ukrainian Strain VI (PVM), the heptapeptide 29AADFEGK35 was found to be recognised by two PVM specific monoclonal antibodies (MAbs) M6D5 and M9G1. This heptapeptide represents the C-terminal part of tryptic tetradecapeptide 22EARPLPTAADFEGK35 (P14) which was also recognised by the same MAbs. The peptides represented sequences of tryptic (P14), thermolysinic (P7) fragments and three heptapeptides containing alanine substitutions for Asp31 and Glu33, were synthesised to determine the contribution of dicarbonic amino acids in the antigen-antibody interaction. All synthetic heptapeptides were recognised by both MAbs weakly in indirect ELISA. These peptides were also used as inhibitors of MAb-CP and MAb-P14 reactions in inhibition ELISA. The results of inhibition ELISA have shown the following: 1) the same concentrations of peptides were more effective to inhibit the interaction of MAbs with P14 than with CP; 2) substitutions of charged amino acids decreased noticeably the ability of peptides to inhibit the antigen-antibody interaction, especially the substitution of Asp31 ; 3) heptapeptides containing alanine substitutions suppressed more effectively the interaction of MAb M6D5 with antigens and were less effective to inhibit the reaction of MAb M9G1 with the same antigens. Thus, the difference in Asp31 and Glu33 contributions to the antigen-antibody complex formation has been found.
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References

[1] Viter SS, Tkachenko TYu, Kolomietz LP, Radavsky YuL. Analysis of antigenic structure of Potato Virus M Ukrainian strains. Biopolym Cell. 2000; 16(4):312-9.
[2] Nikolayeva OV, Novikov VK, Atabekov IG, Kaftanova AS. Changes in the antigenic properties of the potato virus M with progressive degradation of structural protein during isolation and storage. Sel'skokhozyaystvennaya biologiya. 1985; 10:75-8.
[3] Tavantzis SM. Physicochemical properties of potato virus M. Virology. 1984;133(2):427-30.
[4] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[5] Wu GJ, Bruening G. Two proteins from cowpea mosaic virus. Virology. 1971;46(3):596-612.
[6] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
[7] Barany G, Merrifield RB. Solid-phase peptide synthesis. Peptides. Eds E. Gross, J. Meienhoter. New York: Acad, press, 1980. Vol. 2: 1-284.
[8] 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.
[9] Zavriev SK, Kanyuka KV, Levay KE. The genome organization of potato virus M RNA. J Gen Virol. 1991;72 ( Pt 1):9-14.
[10] 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.
[11] 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.
[12] Rajashankar KR, Ramakumar S. Pi-turns in proteins and peptides: Classification, conformation, occurrence, hydration and sequence. Protein Sci. 1996;5(5):932-46.
[13] Benjamin DC, Berzofsky JA, East IJ, Gurd FR, Hannum C, Leach SJ, Margoliash E, Michael JG, Miller A, Prager EM, et al. The antigenic structure of proteins: a reappraisal. Annu Rev Immunol. 1984;2:67-101.
[14] Al Moudallal Z, Briand JP, Van Regenmortel MH. Monoclonal antibodies as probes of the antigenic structure of tobacco mosaic virus. EMBO J. 1982;1(8):1005-10.
[15] Blondel B, Crainic R, Fichot O, Dufraisse G, Candrea A, Diamond D, Girard M, Horaud F. Mutations conferring resistance to neutralization with monoclonal antibodies in type 1 poliovirus can be located outside or inside the antibody-binding site. J Virol. 1986;57(1):81-90.
[16] Collawn JF, Wallace CJ, Proudfoot AE, Paterson Y. Monoclonal antibodies as probes of conformational changes in protein-engineered cytochrome c. J Biol Chem. 1988;263(18):8625-34.