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Overexpression, purification, and functional analysis of recombinant human tubulin dimer
Author(s) -
Minoura Itsushi,
Hachikubo You,
Yamakita Yoshihiko,
Takazaki Hiroko,
Ayukawa Rie,
Uchimura Seiichi,
Muto Etsuko
Publication year - 2013
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2013.08.032
Subject(s) - tubulin , recombinant dna , microtubule , dimer , microbiology and biotechnology , in vitro , function (biology) , cell culture , biology , microtubule associated protein , chemistry , biophysics , biochemistry , genetics , gene , organic chemistry
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 5 mg 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.