Toxic isolectins from the mushroom Boletus venenatus
Toxic isolectins from the mushroom Boletus venenatus
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DOI：
10.1016/j.phytochem.2009.12.003
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发表时间：
2010-04-01
期刊：
影响因子：
3.8
通讯作者：
Kawagishi, Hirokazu
中科院分区：
文献类型：
--
作者：
Horibe, Masashi;Kobayashi, Yuka;Kawagishi, Hirokazu
摄入有毒蘑菇牛肝菌会导致严重的胃肠道综合征，如恶心、反复呕吐、腹泻和胃痛。一个异凝集素家族（B.通过连续80%硫酸铵沉淀、Super Q阴离子交换层析和TSK-凝胶G3000 SW凝胶过滤，从蘑菇中分离出毒性成分venenatus凝集素（BVL）。虽然BVL在SDS-PAGE上显示单一条带，但通过Bio-Assist Q阴离子交换层析，它们进一步被分成8个isolectins（BVL-1至-8）。基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析表明，所有的isolectins显示凝集素活性，并具有非常相似的分子量。其中，BVL-1和BVL-3进一步表征，测定了它们的99个氨基酸的完整氨基酸序列，发现它们彼此相同。在血凝抑制试验中，这两种蛋白质未能结合到任何单或寡核苷酸测试，并显示出相同的糖结合特异性的糖蛋白。在检查的糖蛋白中，去唾液酸胎球蛋白是最强的抑制剂。通过正面亲和层析和表面等离子体共振分析，也分析了每种同工凝集素的糖结合特异性，表明它们识别N-连接的糖链，特别是N-连接的糖链中的Gal β 1-> 4GlcNAc β 1-> 4 Man β 1 -> 4Glc-NAc β 1 -> 4GlcNAc（II型）残基。BVL摄入导致小鼠腹腔内给药后出现致死性毒性，大鼠经口给药后出现腹泻。(C)2009爱思唯尔有限公司版权所有。
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 Bio-Assist 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 Gal beta 1 -> 4GlcNAc beta 1 -> 4Man beta 1 -> 4Glc-NAc beta 1 -> 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. (C) 2009 Elsevier Ltd. All rights reserved.