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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.

Authors
  • Ueda, Minoru
  • Manabe, Yoshiyuki
  • Otsuka, Yuki
  • Kanzawa, Nobuyuki
Type
Published Article
Journal
Chemistry - An Asian Journal
Publisher
Wiley (John Wiley & Sons)
Publication Date
Dec 02, 2011
Volume
6
Issue
12
Pages
3286–3297
Identifiers
DOI: 10.1002/asia.201100392
PMID: 21815269
Source
Medline
License
Unknown

Abstract

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.

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