Toxic isolectins from the mushroom Boletus venenatus.
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概要
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Ingestion of the toxic mushroom Boletus venenatus causes a severe gastrointestinal syndrome, such as nausea, repetitive vomiting, diarrhea, and stomachache. A family of isolectins (B. venenatus lectins, BVLs) was isolated as the toxic principles from the mushroom by successive 80% ammonium sulfate-precipitation, Super Q anion-exchange chromatography, and TSK-gel G3000SW gel filtration. Although BVLs showed a single band on SDS-PAGE, they were further divided into eight isolectins (BVL-1 to -8) by BioAssist Q anion-exchange chromatography. All the isolectins showed lectin activity and had very similar molecular weights as detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. Among them, BVL-1 and -3 were further characterized with their complete amino acid sequences of 99 amino acids determined and found to be identical to each other. In the hemagglutination inhibition assay, both proteins failed to bind to any mono- or oligo-saccharides tested and showed the same sugar-binding specificity to glycoproteins. Among the glycoproteins examined, asialo-fetuin was the strongest inhibitor. The sugar-binding specificity of each isolectin was also analyzed by using frontal affinity chromatography and surface plasmon resonance analysis, indicating that they recognized N-linked sugar chains, especially Galbeta1-->4GlcNAcbeta1-->4Manbeta1-->4GlcNAcbeta1-->4GlcNAc (Type II) residues in N-linked sugar chains. BVLs ingestion resulted in fatal toxicity in mice upon intraperitoneal administration and caused diarrhea upon oral administration in rats.
著者
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中村 祥子
産総研・糖鎖工学セ
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Murata Takeomi
Department Of Applied Biological Chemistry Faculty Of Agriculture Shizuoka University
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Murata Takeomi
Department Of Biological Chemistry Faculty Of Agriculture Shizuoka University
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Kawagishi Hirokazu
Graduate School Of Sci. And Technol. Shizuoka Univ.
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Nakamura Tsuruta
Lectin Application And Analysis Team Research Center For Medical Glycoscience National Institute Of
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Nakamura Sachiko
Glycostructure Analysis Team Research Center For Glycoscience National Institute Of Advanced Industr
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Nakamura Tsuruta
Venture Business Laboratory
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Nakamura Tsuruta
Glycostructure Analysis Team Research Center For Glycoscience National Institute Of Advanced Industr
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Morita Tatsuya
岐阜大学 連合農学研究科
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Nakamura Tsuruta
Research Center For Glycoscience National Institute Of Advanced Industrial Science And Technology (a
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Kobayashi Yuka
Dep. Of Molecular Biotechnology Graduate School Of Advanced Sciences Of Matter Hiroshima Univ.
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Usui Taichi
Department Of Bioscience Graduate School Of Science And Technology Shizuoka University
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