FLAG tagging by CuAAC and nanogram-scale purification of the target protein for a bioactive metabolite involved in circadian rhythmic leaf movement in Leguminosae.
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 x 10(5) differentiated cells. Using this method, we isolated and identified MetE as a cytosolic target protein of potassium isolespedezate, a metabolite controlling plant nyctinasty.
Manabe
Yoshiyuki
Y
Department of Chemistry, Tohoku University, 6-3 Aramakiaza-aoba, Aoba-ku, Sendai 980-8578, Japan.
Mukai
Makoto
M
Ito
Satoko
S
Kato
Nobuki
N
Ueda
Minoru
M
D000480:Alkynes / Q000737:chemistry; D000595:Amino Acid Sequence; D001386:Azides / Q000737:chemistry; D002940:Circadian Rhythm; D007887:Fabaceae / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D000480:Alkynes; D001386:Azides; D010940:Plant Proteins