Molecular and Cellular Biochemistry

, Volume 144, Issue 2, pp 109–116 | Cite as

A 205 kDa protein from non-neuronal cells in culture contains tubulin binding epitopes

  • Clarisa Vial
  • Rosario Armas-Portela
  • Jesús Avila
  • Mauricio González
  • Ricardo B. Maccioni


Microtubule-associated proteins (MAPs) interact with tubulinin vitro andin vivo. Despite that there is a large amount of information on the roles of these proteins in neurons, the data on non-neuronal MAPs or MAPs-related proteins is scarce. There is an increasing number of microtubule-interacting proteins that have been identified in different cultured cell lines, and some of them share common functional epitopes with the most well-known MAPs, MAP-2 and tau. In a search for tubulin-interacting proteins in non-neuronal cells we identified a 205 kDa protein in the monkey kidney Vero cells in culture, on the basis of immunological studies and affinity chromatography. This protein interacts with the C-terminal moiety of β-tubulin and cosediments with taxol assembled microtubules, but it was not recovered after successive cycles of assembly and disassembly. The presence of neuronal MAPs such as MAP-1, MAP-2 and tau was not detected in these cells. Interestingly, the studies showed that the 205 kDa protein contained a tubulin binding motif which was recognized by site-directed antibodies that also tag tubulin binding epitopes on MAP-2 and tau. This characteristic led us to designate this protein as MBD-205, a component that shares binding domains with these MAPs, rather than as a marker of the MAPs family. On the other hand, immunofluorescence experiments using site-specific antibodies, i.e. MAP-reacting monoclonal anti-idiotypic reagent MTB6.22 and a polyclonal antibody to the second tau repeat, revealed a MBD-205 co-localization with membrane structures and microtubule-organizing centers in Vero cells. Microinjection studies along with studies on the cell distribution suggest that MBD-205 appears to play a structural role at the level of the microtubule interactions in these cells.

Key words

MAPs 205 kDa protein Vero cells immunofluorescence cell microinjection analysis 


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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Clarisa Vial
    • 1
  • Rosario Armas-Portela
    • 2
  • Jesús Avila
    • 2
  • Mauricio González
    • 1
  • Ricardo B. Maccioni
    • 1
    • 3
  1. 1.International Center for Cancer and Developmental Biology (ICC)Santiago 7Chile
  2. 2.Centro de Biología Molecular ‘Severo Ochoa’SantiagoChile
  3. 3.Department of Biology, Faculty of SciencesUniversity of ChileSantiagoChile

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