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; Makoto Mukai; Satoko Ito; Nobuki Kato; Minoru Ueda

doi:10.1039/B915843J

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Chemical Communications

Issue 3, 2010

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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

Yoshiyuki Manabe , Makoto Mukai , Satoko Ito , Nobuki Kato and Minoru Ueda

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

DOI: 10.1039/B915843J
Received 03 Aug 2009, Accepted 23 Oct 2009
First published on the web 16 Nov 2009

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Abstract
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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.

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Experimental procedures and spectroscopic data includingcopies of ¹H and ¹³C NMR spectra
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Yoshiyuki Manabe
Makoto Mukai
Satoko Ito
Nobuki Kato
Minoru Ueda

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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

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