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Planta

, Volume 119, Issue 2, pp 113–124 | Cite as

Sieve-tube proteins from Cucurbita maxima

  • J. Beyenbach
  • C. Weber
  • H. Kleinig
Article

Summary

The two main proteins from the phloem exudate of Cucurbita maxima Duchesne have been isolated by ammonium-sulfate precipitation, DEAE-cellulose chromatography, and gel filtration, and have been characterized. They comprise about 40% each of the total protein. The amino-acid composition of these two proteins has been determined. Both are highly basic with an IEP above 9.5. The smaller protein has a molecular weight of ca. 30000 as determined by analytical ultracentrifugation, gel filtration, and SDS polyacrylamide gel electrophoresis. It easily dimerizes to a form which appears to be the naturally occurring structure. The larger protein has a molecular weight of 116000 (ultracentrifugation, SDS polyacrylamide gel electrophoresis). This protein forms a gel in the absence of SH-protecting agents. Several minor proteins have been detected by polyacrylamide gel electrophoresis.

Keywords

Precipitation Molecular Weight Chromatography Total Protein Large Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Becker, D., Kluge, M., Ziegler, H.: Der Einbau von 32PO4 3− in organische Verbindungen durch Siebröhrensaft. Planta (Berl.) 99, 154–162 (1971)Google Scholar
  2. Behnke, H. D., Dörr, I.: Zur Herkunft und Struktur der Plasmafilamente in Assimilatleitbahnen. Planta (Berl.) 74, 18–44 (1967)Google Scholar
  3. Bieleski, R. L.: Phosphorus compounds in translocating phloem. Plant Physiol. 44, 497–502 (1969)Google Scholar
  4. Bryan, J., Wilson, L.: Are cytoplasmic microtubules heteropolymers? Proc. nat. Acad. Sci. (Wash.) 68, 1762–1766 (1971)Google Scholar
  5. Cronshaw, J., Esau, K.: Tubular and fibrillar components of mature and differentiating sieve elements. J. Cell Biol. 34, 801–815 (1967)Google Scholar
  6. Cronshaw, J., Esau, K.: Protein in the phloem of Cucurbita I. The development of protein bodies. J. Cell Biol. 38, 25–39 (1968a).Google Scholar
  7. Cronshaw, J., Esau, K.: P protein in the phloem of Cucurbita II. The P protein of mature sieve elements. J. Cell Biol. 38, 292–303 (1968b)Google Scholar
  8. Eibl, H., Lands, W. E. M.: A new, sensitive determination of phosphate. Analyt. Biochem. 30, 51–57 (1969)Google Scholar
  9. Edelhoch, H.: Spectroscopic determination of tryptophan and tyrosin in proteins. Biochemistry 6, 1948–1954 (1967)Google Scholar
  10. Elias, H. G.: Ultrazentrifugen-Methoden. Munich: Beckman Instruments 1969Google Scholar
  11. Eschrich, W., Evert, R. F., Heyser, W.: Proteins of the sieve tube exudate of Cucurbita maxima. Planta (Berl.) 100, 208–221 (1971)Google Scholar
  12. Feit, H., Slusarek, L., Shelanski, M. L.: Heterogeneity of tubulin subunits. Proc. nat. Acad. Sci. (Wash.) 68, 2028–2031 (1971)Google Scholar
  13. Fine, R. E.: Heterogeneity of tubulin. Nature (Lond.) New Biol. 233, 283–284 (1971)Google Scholar
  14. Gardner, D. C. J., Peel, A. J.: ATP in sieve tube sap from willow. Nature (Lond.) 222, 747 (1969)Google Scholar
  15. Gilder, J., Cronshaw, J.: ATPase in the phloem of Cucurbita. Planta (Berl.) 110, 189–204 (1973)Google Scholar
  16. Gilder, J., Cronshaw, J.: A biochemical and cytochemical study of ATPase activity in the phloem of Nicotiana tabacum. J. Cell Biol. 60, 221–235 (1974)Google Scholar
  17. Hart, J. W., Sabnis, D. D.: Colchicine-binding protein from phloem and xylem of a higher plant. Planta (Berl.) 109, 147–152 (1973)Google Scholar
  18. Kleinig, H., Dörr, I., Weber, C., Kollmann, R.: Filamentous proteins from plant sieve tubes. Nature (Lond.) New Biol. 229, 152–153 (1971a)Google Scholar
  19. Kleinig, H., Dörr, I., Kollmann, R.: Vinblastine-induced precipitation of phloem proteins in vitro. Protoplasma (Wien) 73, 293–302 (1971b)Google Scholar
  20. Kluge, M., Becker, D., Ziegler, H.: Untersuchungen über ATP und andere organische Phosphorverbindungen im Siebröhrensaft von Yucca flaccida und Salix triandra. Planta (Berl.) 91, 68–79 (1970)Google Scholar
  21. Kollmann, R., Dörr, I., Kleinig, H.: Protein filaments—structural components of the phloem exudate. I. Observations with Cucurbita and Nicotiana. Planta (Berl.) 95, 86–94 (1970)Google Scholar
  22. Lehmann, J.: Zur Lokalisation von Dehydrogenasen des Energiestoffwechsels im phloem von Cucurbita pepo L. Planta (Berl.) 111, 187–198 (1973a)Google Scholar
  23. Lehmann, J.: Untersuchungen am Phloemexudat von Cucurbita pepo L. I. Enzymaktivitäten von Glykolyse, Gärung und Citrat-Zyklus. Planta (Berl.) 114, 41–51 (1973b)Google Scholar
  24. Lehmann, J.: Untersuchungen am Phloemexudat von Cucurbita pepo L. II. Enzymaktivitäten der Gluconeogenese und des Auf- und Abbaus von Di- und Polysacchariden. Planta (Berl.) 114, 51–61 (1973c)Google Scholar
  25. Lowry, D. H., Rosebrough, N. J., Farr, N. L., Randall, R. L.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)Google Scholar
  26. Parthasarathy, M. V., Mühlethaler, K.: Ultrastructure of protein tubules in differentiating sieve elements. Cytobiol. 1, 17–36 (1969)Google Scholar
  27. Pollard, T. D., Korn, E. D.: Acanthamoeba myosin. I. Isolation from Acanthamoeba castellanii of an enzyme similar to muscle myosin. J. biol. Chem. 248, 4682–8690 (1973)Google Scholar
  28. Shapiro, A. L., Vinuela, E., Maizel, J. V.: Molecular-weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochim. biophys. Res. Commun. 28, 815–820 (1967)Google Scholar
  29. Spencer, R. L., Wold, F.: A new convenient method for estimation of total cysteinecystine in proteins. Analyt. Biochem. 32, 185–190 (1969)Google Scholar
  30. Tuppy, H., Wiesbauer, T., Wintersberger, E.: Aminosäure-p-nitroanilide als Substrate für Aminopeptidasen und andere proteolytische Fermente. Hoppe-Seylers Z. physiol. Chem. 329, 278–288 (1962)Google Scholar
  31. Walker, T. S.: The purification and some properties of a protein causing gelling in phloem sieve tube exudate from Cucurbita pepo. Biochim. biophys. Acta (Amst.) 257, 433–444 (1972)Google Scholar
  32. Walker, T. S., Thaine, R.: Proteins and fine structural components in exudate from sieve tubes in Cucurbita pepo stems. Ann. Bot. 35, 773–790 (1971)Google Scholar
  33. Weber, C., Kleinig, H.: Molecular weights of Cucurbita sieve tube proteins. Planta (Berl.) 99, 179–182 (1971)Google Scholar
  34. Yapa, P. A. J., Spanner, D. C.: Isoelectric focusing of sieve tube protein. Planta (Berl.) 106, 369–373 (1972)Google Scholar
  35. Zacharius, R. M., Zell, T. E., Morrison, J. H., Woodlock, J. J.: Glycoprotein staining following electrophoresis on acrylamide gels. Analyt. Biochem. 30, 148–152 (1969)Google Scholar

Copyright information

© Springer-Verlag 1974

Authors and Affiliations

  • J. Beyenbach
    • 1
  • C. Weber
    • 1
  • H. Kleinig
    • 1
  1. 1.Institut für Biologie II, Lehrstugl für ZellbiologieUniversität FreiburgFreiburg i.Br.Federal Republic of Germany

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