Development of biotin-avidin technology to investigate okadaic acid-promoted cell signaling pathway

Keiichi Konoki, Naoyuki Sugiyama, Michio Murata, Kazuo Tachibana, Yasumaru Hatanaka

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

Four biotin conjugates of okadaic acid were synthesized for evaluating their interactions with protein phosphatase 2A (PP2A) by surface plasmon resonance (SPR). C7-biotinylated okadaic acid exhibited the strongest binding affinity to the enzyme, while Cl-biotinylated derivative was devoid of affinity. C24- or C27-biotinylated okadaic acid showed moderate affinity to the enzyme. In the wake of this finding, a biotinyl photoaffinity probe was introduced into 7-OH of okadaic acid. Photoaffinity labeling followed by SDS-PAGE analysis indicated that the okadaic acid derivative clearly labeled PP2A. Furthermore, three proteins were also labeled in crude extracts of a marine sponge Halichondria okadai. All these results imply that the C7-biotin conjugate is a versatile reagent for biochemical studies of okadaic acid-binding proteins including PP2A. (C) 2000 Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)9003-9014
Number of pages12
JournalTetrahedron
Volume56
Issue number46
DOIs
Publication statusPublished - 2000 Nov 10

Keywords

  • Okadaic acid
  • Photoaffinity labeling
  • Protein phosphatase
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Organic Chemistry

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