1887

Abstract

SUMMARY

The b protein, produced in leaves of cv. Xanthi-nc following infection with tobacco mosaic virus, has been purified to homogeneity by a procedure which involves gel chromatography and absorption on to DEAE-cellulose. One gel chromatography step was sufficient when the procedure was applied to leaf extracts made in an acid buffer, whereas two were necessary with extracts made at pH 8. The final product migrates as a single protein band on electrophoresis in both acrylamide and SDS-acrylamide gels. Its mol. wt. is estimated to be 15000 by electrophoresis and 14200 by ultracentrifugation. Amino acid analysis suggests that it contains about 136 residues of which 39 are potentially acidic, 13 basic and 16 aromatic. The absorbance coefficient is estimated to be 18.9. No evidence was found for the presence of a nucleotide component.

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1978-05-01
2024-12-06
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References

  1. Babul J., Stellwagen E. 1969; Measurement of protein concentration with interference optics. Analytical Biochemistry 28:216–221
    [Google Scholar]
  2. Bencze W. L., Schmid K. 1957; Determination of tyrosine and tryptophan in proteins. Analytical Chemistry 29:1193–1196
    [Google Scholar]
  3. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  4. Cohen P. 1973; The subunit structure of rabbit-skeletal-muscle phosphorylase kinase, and the molecular basis of its activation reactions. European Journal of Biochemistry 34:1–14
    [Google Scholar]
  5. Cohn E. J., Edsall J. T. 1943 In Proteins, Amino Acids and Peptides New York: Rheinhold Publishing Corporation;
    [Google Scholar]
  6. Davis B. J. 1964; Disc electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121:404–427
    [Google Scholar]
  7. Gianinazzi S., Kassanis B. 1974; Virus resistance induced in plants by polyacrylic acid. Journal of General Virology 23:1–9
    [Google Scholar]
  8. Gianinazzi S., Pratt H. M., Shewry P. R., Miflin B. J. 1977; Partial purification and preliminary characterization of soluble leaf proteins specific to virus infected tobacco plants. Journal of General Virology 34:345–351
    [Google Scholar]
  9. Gianinazzi S., Vallee J. C., Martin C. 1969; Hypersensibilite aux virus, temperature et proteines soluble chez le Nicotiana Xanthi n.c. Compte rendu de l’Academic des Sciences de Paris 268:800–802
    [Google Scholar]
  10. Holbrook L. B., Leaver A. G. 1976; A procedure to increase the sensitivity of staining by Coomassie brilliant blue G250-perchloric acid solution. Analytical Biochemistry 75:634–636
    [Google Scholar]
  11. Israel H. W., Ross A. F. 1967; The fine structure of local lesions induced by TMV in tobacco. Virology 33:272–286
    [Google Scholar]
  12. Kassanis B., Gianinazzi S., White R. F. 1974; A possible explanation of the resistance of virus-infected tobacco plants to second infection. Journal of General Virology 23:11–16
    [Google Scholar]
  13. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  14. Martin C., Gallet M. 1966; Nouvelle observations sur le phenomene d’hypersensibilite aux virus chez vegetaux. Compte rendu de I’Academie des Sciences de Paris 263:1316–1318
    [Google Scholar]
  15. Ornstein L. 1964; Disc electrophoresis. I. Background and theory. Annals of the New York Academy of Sciences 121:321–349
    [Google Scholar]
  16. Reisner A. H., Nemes P., Bucholtz C. 1975; The use of Coomassie brilliant blue G250 perchloric acid solution for staining in electrophoresis and isoelectric focussing on polyacrylamide gels. Analytical Biochemistry 64:509–516
    [Google Scholar]
  17. Rohloff H., Lerch B. 1977; Soluble leaf proteins in virus infected plants and acquired resistance. I. Investigations on Nicotiana tabacum cvs. ‘Xanthi-nc’ and ‘Samsun’. Phytopathologische Zeitschrift 89:306–316
    [Google Scholar]
  18. Ross A. F. 1961a; Localized acquired resistance to plant virus infection in hypersensitive hosts. Virology 14:329–339
    [Google Scholar]
  19. Ross A. F. 1961b; Systemic acquired resistance induced by localised virus infections in plants. Virology 14:340–358
    [Google Scholar]
  20. Tas P. W. L., Peters D. 1977; The occurrence of a soluble protein (E1) in cucumber cotyledons infected with plant viruses. Netherlands Journal of Plant Pathology 83:5–12
    [Google Scholar]
  21. Van Loon L. C. 1975; Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var. ‘Samsun’ and ‘Samsun NN’ III. Influence of temperature and virus strain on changes induced by tobacco mosaic virus. Physiological Plant Pathology 6:289–300
    [Google Scholar]
  22. Van Loon L. C., Van Kammen A. 1970; Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var. ‘Samsun’ and ‘Samsun NN’. II. Changes in protein constitution after infection with tobacco mosaic virus. Virology 40:199–211
    [Google Scholar]
  23. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulphate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
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