Detection of Rap1A as a yessotoxin binding protein from blood cell membranes

Satoru Ujihara, Tohru Oishi, Ryota Mouri, Rie Tamate, Keiichi Konoki, Nobuaki Matsumori, Michio Murata, Yasukatsu Oshima, Naoyuki Sugiyama, Masaru Tomita, Yasushi Ishihama

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10 Citations (Scopus)

Abstract

As is the case with other ladder-shaped polyether compounds, yessotoxin is produced by marine dinoflagellate, and possesses various biological activities beside potent toxicity. To gain a better understanding of the molecular mechanism for high affinity between these polyethers and their binding proteins, which accounts for their powerful biological activities, we searched for its binding proteins from human blood cells by using the biotin-conjugate of desulfated YTX as a ligand. By a protein pull-down protocol with use of streptavidin beads, a band of specifically binding proteins was detected in SDS-PAGE. HPLC-tandem mass spectrometry (MS/MS) indicated that Rap 1A, one of Ras superfamily proteins, binds to the YTX-linked resins. Western blotting and surface plasmon resonance experiments further confirmed that Rap1A specifically binds to YTX with the KD value around 4 μM.

Original languageEnglish
Pages (from-to)6443-6446
Number of pages4
JournalBioorganic and Medicinal Chemistry Letters
Volume20
Issue number22
DOIs
Publication statusPublished - 2010 Nov 15

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Keywords

  • Ladder-shaped polyether
  • Pull-down assay
  • Ras superfamily proteins
  • Yessotoxin
  • ap1A

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Ujihara, S., Oishi, T., Mouri, R., Tamate, R., Konoki, K., Matsumori, N., Murata, M., Oshima, Y., Sugiyama, N., Tomita, M., & Ishihama, Y. (2010). Detection of Rap1A as a yessotoxin binding protein from blood cell membranes. Bioorganic and Medicinal Chemistry Letters, 20(22), 6443-6446. https://doi.org/10.1016/j.bmcl.2010.09.080