A novel tubulin from Mimosa pudica. Purification and characterization

Eur J Biochem. 1990 Sep 11;192(2):329-35. doi: 10.1111/j.1432-1033.1990.tb19231.x.

Abstract

From Mimosa pudica fresh leaves and pulvinar callus cells, we have purified tubulin protein using an anion-exchange resin, DEAE-Sephadex A-50, followed by ammonium sulfate fractionation and Sephadex G-200 gel filtration. The purified protein consisted of alpha and beta subunits and trace quantities of other proteins. When analysed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis at pH 8.8, both alpha and beta subunits of tubulin almost comigrated with their counterparts from goat brain. Two-dimensional electrophoresis revealed that this tubulin contains one major alpha-tubulin having a pI value of 7.1 and three beta species having pI values of 6.70, 6.46 and 6.40. Morphologically normal microtubules were observed by electron microscopy; self-polymerization in vitro, even in the cold, can also be achieved. Radioimmunoassays, and also immunoblotting with the antibodies raised against alpha- and beta-tubulins of this plant, showed that the nature of alpha-tubulin is different from that obtained from other sources. This is an example of plant tubulin where strong colchicine binding at 1 microM was observed. This protein constitutes 5-6% of the total extractable protein in the leaves. We propose that movement of the leaves of this plant may be regulated by the presence of a high amount of this protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Macromolecular Substances
  • Microscopy, Electron
  • Molecular Weight
  • Peptide Mapping
  • Plants / metabolism*
  • Tubulin / isolation & purification*
  • Tubulin / metabolism
  • Tubulin / ultrastructure

Substances

  • Macromolecular Substances
  • Tubulin