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Cassia obtusifolia MetE as a Cytosolic Target for Potassium Isolespedezate, a Leaf‐Opening Factor of Cassia plants: Target Exploration by a Compact Molecular‐Probe Strategy
Author(s) -
Ueda Minoru,
Manabe Yoshiyuki,
Otsuka Yuki,
Kanzawa Nobuyuki
Publication year - 2011
Publication title -
chemistry – an asian journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 106
eISSN - 1861-471X
pISSN - 1861-4728
DOI - 10.1002/asia.201100392
Subject(s) - cassia , ligand (biochemistry) , affinity chromatography , chemistry , target protein , cytosol , combinatorial chemistry , chromatography , biochemistry , receptor , enzyme , medicine , alternative medicine , pathology , traditional chinese medicine , gene
Affinity chromatography by using ligand‐immobilized bead technology is generally the first choice for target exploration of a bioactive ligand. However, when a ligand has comparatively low affinity against its target, serious difficulties will be raised in affinity‐based target detection. We report here that the use of compact molecular probes (CMP) will be advantageous in such cases; it enables the retention of moderate affinity between the ligand and its target in contrast to immobilizing the ligand on affinity beads that will cause a serious drop in affinity to preclude target detection. In the CMP strategy, a CMP containing an azide handle is used for an initial affinity‐based labeling of target, and subsequent tagging by CuAAC with a large FLAG tag will give a tagged target protein. By using the CMP strategy, we succeeded in the identification of Cassia obtusifolia MetE as a cytosolic target protein of potassium isolespedezate ( 1 ), a moderately bioactive ligand.