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. 1985 Oct;4(10):2451–2455. doi: 10.1002/j.1460-2075.1985.tb03955.x

Identification of multiple tubulins in taxol microtubules purified from carrot suspension cells

PJ Dawson 1, CW Lloyd 1
PMCID: PMC554528  PMID: 15929221

Abstract

Tubulin has been purified from carrot suspension cells by ionexchange chromatography and assembled into microtubules in the presence of 20 μM taxol. One-dimensional SDS-PAGE suggested that the α band migrated faster than the β band (as has been established for some lower eukaryotic tubulins) and this heterology with brain tubulins was confirmed by peptide mapping. When subjected to two-dimensional gel electrophoresis, the plant tubulins could be separated into multiple α and β isotypes. Immunoblotting, using monoclonal antitubulins, confirmed that the tubulin isotypes identified in taxol microtubules represent all of the tubulins present in homogenates of unsynchronised log-phase carrot suspension cells. All identified tubulins are therefore assembly-competent under these conditions. Plant cells can contain four different microtubule arrays, but cells arrested in G0/G1 contain only cortical microtubule arrays; such cells, however, exhibit the same tubulin profile as non-synchronised cells, thereby showing no restriction in the number of subunits during this phase of the cell cycle.

Keywords: tubulin, taxol, microtubules, plant suspension cells

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