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The association of carbohydrate with tubulin and in vitro assembled microtubules from bovine brain

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Phosphocellulose-purified tubulin (PC tubulin) was analyzed for neutral and amino sugar content, which was found to be 8.3±0.11 and 0.8±0.02 mol/mol dimer, respectively. A histochemical-electron-microscopic investigation was undertaken to attempt to localize carbohydrate associated with polymerized microtubules (MT). Outer diameters of MT assembled in vitro from bovine brain MT protein (tubulin and microtubule associated proteins) were found to increase upon treatment with ruthenium red, Alcian blue, and lanthanum hydroxide, which have been reported to possess specificity for complex carbohydrates. Concanavalin A-reactive sites were detected on the surface and in the lumen of MT assembled from MT protein and from PC tubulin.

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References

  • Behnke, O.: Studies on isolated microtubules. Evidence for a clear space component. Cytobiologie 11, 366–381 (1975)

    Google Scholar 

  • Bernhard, W., Avrameas, S.: Ultrastructural visualization of cellular carbohydrate components by means of Concanavalin A. Exp. Cell Res. 64, 232–236 (1971)

    Google Scholar 

  • Borisy, G.G., Olmsted, J.B., Marcum, J.M., Allen, C.: Microtubule, assembly in vitro. Fed. Proc. 33, 167–174 (1974)

    Google Scholar 

  • Burton, P.R., Fernandez, H.L.: Delineation by lanthanum staining of filamentous elements associated with the surfaces of axonal microtubules. J. Cell. Sci. 12, 567–583 (1973)

    Google Scholar 

  • Burton, P.R., Hinkley, R.E.: Further electron microscopic characterization of axoplasmic microtubules of the ventral nerve cord of the crayfish. J. Submicrose. Cytol. 6, 311–326 (1974)

    Google Scholar 

  • Eipper, B.A.: Rat brain microtubule protein: purification and determination of covalently bound phosphate and carbohydrate. Proc. Natl. Acad. Sci. USA 69, 2283–2287 (1972)

    Google Scholar 

  • Falxa, M.L., Gill, T.J.: Preparation and properties of an alkylated brain protein related to the structural subunit of microtubules. Arch. Biochem. Biophys. 135, 194–200 (1969)

    Google Scholar 

  • Feit, H., Shelanski, M.L.: Is tubulin a glycoprotein? Biochem. Biophys. Res. Commun. 66, 920–927 (1975)

    Google Scholar 

  • Goodman, D.B.P., Rasmussen, H., DiBella, F., Guthrow, C.E.: Cyclic adenosine 3′: 5′-monophosphate-stimulated phosphorylation of isolated neurotubule subunits. Proc. Natl. Acad. Sci. USA 67, 652–659 (1970)

    Google Scholar 

  • Graham, R.C., Karnovsky, M.J.: The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J. Histochem. Cytochem. 14, 291–302 (1966)

    Google Scholar 

  • Hinkley, R.E.: Axonal microtubules and associated filaments stained by Alcian blue. J. Cell Sci. 13, 753–761 (1973)

    Google Scholar 

  • Hüttich, V.K., Müller, H., Unger, E.: Concanavalin A-Bindung an zyklisch reassemblierte Mikrotubuli aus Schweinehirn-Tubulin. Acta Histochem. (Jena) 58, 324–331 (1977)

    Google Scholar 

  • Karnovsky, M.J.: A formaldehyde-glutaraldehyde fixative of high osmolatity for use in electron microscopy. J. Cell Biol. 27, 137–138 (1965)

    Google Scholar 

  • Krystal, G., Graham, A.F.: A sensitive method for estimating the carbohydrate content of glycoproteins. Anal. Biochem. 70, 336–345 (1976)

    Google Scholar 

  • Laemmli, U.K.: Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970)

    Google Scholar 

  • Lane, N.J., Treherne, J.E.: Lanthanum staining of neurotubules in axons from cockroach ganglia. J. Cell Sci. 7, 217–231 (1970)

    Google Scholar 

  • Larsson, H., Wallin, M., Edström, A.: Induction of a sheet polymer of tubulin by Zn2+. Exp. Cell Res. 100, 104–110 (1976)

    Google Scholar 

  • Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randall, R.J.: Protein measurements with folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Margolis, R.K., Margolis, R.U., Shelanski, M.L.: The carbohydrate content of brain microtubule protein. Biochem. Biophys. Res. Commun. 47, 432–437 (1972)

    Google Scholar 

  • Stebbings, H., Bennet, C.E.: The sleeve element of microtubules. In: Microtubules and microtubule inhibitors (eds. M. Borgers, M. deBrabander) Amsterdam: North-Holland 1975

    Google Scholar 

  • Stephens, R.E.: Major membrane protein differences in cilia, and flagella: evidence for a membraneassociated tubulin. Biochemistry 16, 2047–2058 (1977)

    Google Scholar 

  • Tani, E., Ametani, T.: Extracellular distribution of ruthenium red-positive substance in the cerebral cortex. J. Ultrastruc. Res. 34, 1–14 (1971)

    Google Scholar 

  • Weingarten, M.D., Lockwood, A.H., Hwo, S-Y., Kirshner, M.W.: A protein factor essential for microtubule assembly. Proc. Natl. Acad. Sci. USA 72, 1858–1862 (1975)

    Google Scholar 

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This work was supported by grants from Statens Naturvetenskapliga. Forsknigsråd (No. B0294-020), Stiftelsen Lars Hiertas Minne, Adlerbertska Forskningsfonden, Hierta-Retzius' Stipendiefond, Anna Ahrenbergs Fond, and Wilhelm och Martina Lundgrens Vetenskapsfond. Appreciation is extended to Inger Holmqvist for her excellent technical assistance

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Prus, K., Mattisson, A. The association of carbohydrate with tubulin and in vitro assembled microtubules from bovine brain. Histochemistry 61, 281–289 (1979). https://doi.org/10.1007/BF00508449

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