Isolation of novel prototype galectins from the marine ball sponge Cinachyrella sp. guided by their modulatory activity on mammalian glutamate-gated ion channels.
Isolation of novel prototype galectins from the marine ball sponge Cinachyrella sp. guided by their modulatory activity on mammalian glutamate-gated ion channels.
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从海洋球海绵 Cinachyrella sp 中分离新型原型半乳糖素。
DOI：
10.1093/glycob/cws165
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发表时间：
2013
期刊：
影响因子：
4.3
通讯作者：
Sakai,Ryuichi
中科院分区：
文献类型：
--
作者：
Ueda,Takuya;Nakamura,Yuka;Smith,CalebM;Copits,BryanA;Inoue,Akira;Ojima,Takao;Matsunaga,Satoko;Swanson,GeoffreyT;Sakai,Ryuichi
在这里，我们报道了生物活性引导下从海绵体中分离新的凝集素。，采自日本伊洛摩特岛。我们将凝集素蛋白称为cachyrellagalectin (CchGs)，发现其在海绵水提取物中具有调节哺乳动物嗜离子性谷氨酸受体功能的活性。半乳糖苷比甘露糖更能有效地竞争兔红细胞的CchGs聚集，而甘露糖是低聚糖结合蛋白半乳糖素家族成员的特征。凝集素活性非常稳定，在100°C下暴露1小时后，血凝活性仅略有下降，并且对钙浓度几乎不敏感。CchG-1和cchg -2在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中分别显示为16和18 kDa，而基质辅助激光解吸电离-飞行时间质谱分析显示，CchG-1和cchg -2的中心分别为16,216和16,423。利用Edman降解和cDNA克隆相结合的方法推导出CchGs的氨基酸序列，发现CchGs与动物原型凝集素具有远缘同源关系，并且由多种分离凝集素组成。其中一种分离凝集素以重组蛋白的形式表达，具有与天然凝集素相当的物理化学和生物学特性。CchGs的生化特性以及它们对哺乳动物兴奋性氨基酸受体的意外活性表明，对这些新成员的进一步分析将产生进一步的糖生物学和神经生理学见解。
Here we report the bioactivity-guided isolation of novel galectins from the marine spongeCinachyrellasp., collected from Iriomote Island, Japan. The lectin proteins, which we refer to as theCinachyrellagalectins (CchGs), were identified as the active principles in an aqueous sponge extract that modulated the function of mammalian ionotropic glutamate receptors. Aggregation of rabbit erythrocytes by CchGs was competed most effectively by galactosides but not mannose, a profile characteristic of members of the galectin family of oligosaccharide-binding proteins. The lectin activity was remarkably stable, with only a modest loss in hemagglutination after exposure of the protein to 100°C for 1 h, and showed little sensitivity to calcium concentration. CchG-1 and -2 appeared as 16 and 18 kDa in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, respectively, whereas matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry indicated broad ion clusters centered at 16,216 and 16,423, respectively. The amino acid sequences of the CchGs were deduced using a combination of Edman degradation and cDNA cloning and revealed that the proteins were distant orthologs of animal prototype galectins and that multiple isolectins comprised the CchGs. One of the isolectins was expressed as a recombinant protein and exhibited physico-chemical and biological properties comparable with those of the natural lectins. The biochemical properties of the CchGs as well as their unexpected activity on mammalian excitatory amino acid receptors suggest that further analysis of these new members of the galectin family will yield further glycobiological and neurophysiological insights.