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Journal of Cell Science, Vol 92, Issue 3 519-528, Copyright © 1989 by Company of Biologists
JOURNAL ARTICLES |
P Draber, E Draberova, I Linhartova and V Viklicky
Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Prague.
A panel of 11 monoclonal antibodies specific to alpha- or beta-tubulin subunits was used to study the location of tubulin molecules in cytoplasmic microtubules. Specificity of antibodies was confirmed by immunoblotting and immunofluorescence experiments on fixed cells. The limited proteolysis of tubulin with trypsin and chymotrypsin followed by immunoblotting demonstrated that the antibodies discriminated between structural domains of both subunits. Epitope mapping of isolated alpha-tubulin revealed that a set of antibodies against the N-terminal domain of the alpha-subunit (TU-01, TU-02, TU-03, TU-09, 6-11B-1) recognized at least four different antigenic determinants. Immunofluorescence staining of unfixed detergent-extracted cells showed that antibodies to determinants on C-terminal domains labelled microtubules, but these were not decorated with antibodies to N-terminal domains. The same results were obtained after microinjection of antibodies into living cells. The unchanged distribution of microtubules in injected cells was confirmed by double-label immunofluorescence with polyclonal antibodies. The data indicate that while parts of C-terminal domains of both subunits are exposed on the exterior of the microtubules, considerable regions of the N-terminal domains are either not exposed on the surface of cytoplasmic microtubules, or are masked by interacting proteins.
This article has been cited by other articles:
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  V. Sulimenko, E. Draberova, T. Sulimenko, L. Macurek, V. Richterova, P. Draber, and P. Draber Regulation of Microtubule Formation in Activated Mast Cells by Complexes of {gamma}-Tubulin with Fyn and Syk Kinases. J. Immunol., June 15, 2006; 176(12): 7243 - 7253. [Abstract] [Full Text] [PDF]
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 J. Peknicova, A. Kubatova, V. Sulimenko, E. Draberova, V. Viklicky, P. Hozak, and P. Draber Differential Subcellular Distribution of Tubulin Epitopes in Boar Spermatozoa: Recognition of Class III {beta}-Tubulin Epitope in Sperm Tail Biol Reprod, September 1, 2001; 65(3): 672 - 679. [Abstract] [Full Text] [PDF]
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P. Binarová, V. Cenklová, B. Hause, E. Kubátová, M. Lysák, J. Doleel, L. Bögre, and P. Dráber Nuclear {gamma}-Tubulin during Acentriolar Plant Mitosis PLANT CELL, March 1, 2000; 12(3): 433 - 442. [Abstract] [Full Text]
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 L. Draberova, E. Draberova, Z. Surviladze, P. Draber, and P. Draber Protein tyrosine kinase p53/p56lyn forms complexes with {gamma}-tubulin in rat basophilic leukemia cells Int. Immunol., November 1, 1999; 11(11): 1829 - 1839. [Abstract] [Full Text] [PDF]
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 A. J. Hall, H. L. Vos, and R. M. Bertina Lipopolysaccharide Induction of Tissue Factor in THP-1 Cells Involves Jun Protein Phosphorylation and Nuclear Factor kappa B Nuclear Translocation J. Biol. Chem., January 1, 1999; 274(1): 376 - 383. [Abstract] [Full Text] [PDF]
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 E Draberova and P Draber A microtubule-interacting protein involved in coalignment of vimentin intermediate filaments with microtubules J. Cell Sci., January 12, 1993; 106(4): 1263 - 1273. [Abstract] [PDF]
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