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

Minoru Ueda, Yoshiyuki Manabe, Yuki Otsuka, Nobuyuki Kanzawa

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

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. Affinity chromatography by using ligand-immobilized bead technology is the first choice for target exploration of a bioactive ligand. However, when a ligand has comparatively low affinity against its target, serious difficulties will arise. Herein, we report that the use of a compact molecular probe (CMP) will be advantageous in such cases (see scheme).

Original languageEnglish
Pages (from-to)3286-3297
Number of pages12
JournalChemistry - An Asian Journal
Volume6
Issue number12
DOIs
Publication statusPublished - 2011 Dec 2

Keywords

  • Cassia obtusifolia
  • affinity chromatography
  • molecular probes
  • natural products
  • proteins

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry

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