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

Research output: Contribution to journalArticle

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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 - Nov 15 2010

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Cell membranes
Blood Cells
Carrier Proteins
Blood
Polyethers
Cell Membrane
Bioactivity
Dinoflagellida
ras Proteins
Streptavidin
Surface Plasmon Resonance
Ladders
Surface plasmon resonance
Biotin
Tandem Mass Spectrometry
Mass spectrometry
Toxicity
Polyacrylamide Gel Electrophoresis
Proteins
Resins

All Science Journal Classification (ASJC) codes

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

Cite this

Detection of Rap1A as a yessotoxin binding protein from blood cell membranes. / Ujihara, Satoru; Oishi, Tohru; Mouri, Ryota; Tamate, Rie; Konoki, Keiichi; Matsumori, Nobuaki; Murata, Michio; Oshima, Yasukatsu; Sugiyama, Naoyuki; Tomita, Masaru; Ishihama, Yasushi.

In: Bioorganic and Medicinal Chemistry Letters, Vol. 20, No. 22, 15.11.2010, p. 6443-6446.

Research output: Contribution to journalArticle

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, vol. 20, no. 22, pp. 6443-6446. https://doi.org/10.1016/j.bmcl.2010.09.080
Ujihara, Satoru ; Oishi, Tohru ; Mouri, Ryota ; Tamate, Rie ; Konoki, Keiichi ; Matsumori, Nobuaki ; Murata, Michio ; Oshima, Yasukatsu ; Sugiyama, Naoyuki ; Tomita, Masaru ; Ishihama, Yasushi. / Detection of Rap1A as a yessotoxin binding protein from blood cell membranes. In: Bioorganic and Medicinal Chemistry Letters. 2010 ; Vol. 20, No. 22. pp. 6443-6446.
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