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FEBS Lett. 2013 Nov 1;587(21):3450-5. doi: 10.1016/j.febslet.2013.08.032. Epub 2013 Sep 8.

Overexpression, purification, and functional analysis of recombinant human tubulin dimer.

Author information

1
Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

Abstract

Microtubules consisting of tubulin dimers play essential roles in various cellular functions. Investigating the structure-function relationship of tubulin dimers requires a method to prepare sufficient quantities of recombinant tubulin. To this end, we simultaneously expressed human α1- and β3-tubulin using a baculovirus-insect cell expression system that enabled the purification of 5mg recombinant tubulin per litre of cell culture. The purified recombinant human tubulin could be polymerized into microtubules that glide on a kinesin-coated glass surface. The method provides a powerful tool for in vitro functional analyses of microtubules.

KEYWORDS:

Baculovirus-insect cell expression; Kinesin motility; MAP; PTM; Tubulin assembly; WT; microtubule-associated protein; posttranslational modification; wild-type

PMID:
24021646
DOI:
10.1016/j.febslet.2013.08.032
[Indexed for MEDLINE]
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