Issue 3, 2010
From the journal:

Chemical Communications

FLAG tagging by CuAAC and nanogram-scale purification of the target protein for a bioactive metabolite involved in circadian rhythmic leaf movement in Leguminosae†

Yoshiyuki Manabe,a   Makoto Mukai,a   Satoko Ito,a   Nobuki Katoa  and  Minoru Ueda*a  

* Corresponding authors

a Department of Chemistry, Tohoku University, 6-3 Aramakiaza-aoba, Aoba-ku, Sendai 980-8578, Japan
E-mail: ueda@mail.tains.tohoku.ac.jp
Fax: (+) 81-22-795-6553
Tel: (+) 81-22-795-6553

Abstract

We report a stepwise FLAG-tagging strategy for the purification of target proteins for bioactive metabolites. This method realizes the microscale purification and identification of target protein from as few as 1 × 105 differentiated cells. Using this method, we isolated and identified MetE as a cytosolic target protein of potassium isolespedezate, a metabolite controlling plant nyctinasty.

Graphical abstract: FLAG tagging by CuAAC and nanogram-scale purification of the target protein for a bioactive metabolite involved in circadian rhythmic leaf movement in Leguminosae

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Article information

DOI
https://doi.org/10.1039/B915843J
Article type
Communication
Submitted
03 Aug 2009
Accepted
23 Oct 2009
First published
16 Nov 2009

Chem. Commun., 2010,46, 469-471

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FLAG tagging by CuAAC and nanogram-scale purification of the target protein for a bioactive metabolite involved in circadian rhythmic leaf movement in Leguminosae

Y. Manabe, M. Mukai, S. Ito, N. Kato and M. Ueda, Chem. Commun., 2010, 46, 469 DOI: 10.1039/B915843J

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