Characterization and cloning of GNA-like lectin from the mushroom Marasmius oreades
Characterization and cloning of GNA-like lectin from the mushroom Marasmius oreades
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DOI：
10.1007/s10719-012-9401-6
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发表时间：
2012-06
影响因子：
3
通讯作者：
Michiko Shimokawa;Ayako Fukudome;Ryoko Yamashita;Y. Minami;F. Yagi;H. Tateno;J. Hirabayashi
中科院分区：
文献类型：
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作者：
Michiko Shimokawa;Ayako Fukudome;Ryoko Yamashita;Y. Minami;F. Yagi;H. Tateno;J. Hirabayashi
在无唾液酸胎球蛋白柱上从日本生长的 Marasmius oreades 的子实体中纯化出一种新的甘露糖识别凝集素 (MOL)。众所周知，来自同一真菌子实体的凝集素 (MOA) 是一种识别 B 型血糖的核糖体失活型凝集素。然而，在我们的初步调查中，在日本M.的子实体中并未发现MOA。结果，MOL 被分离出来。凝胶过滤显示 MOL 是与两个 13 kDa 亚基非共价结合的同二聚体。 MOL 的 N 端序列被封闭。 MOL 的序列是通过 cDNA 克隆和酶消化肽的蛋白质测序来确定的。该序列显示了来自植物的球茎型甘露糖结合凝集素的甘露糖结合基序，以及与序列的相似性。通过血凝抑制对糖结合特异性的分析揭示了 MOL 对甘露糖和甲状腺球蛋白的偏好，但脱唾液酸胎球蛋白是测试的最强的糖蛋白抑制剂。此外，聚糖阵列分析表明，MOL 的特异性模式与典型的甘露糖特异性凝集素不同。 MOL 更喜欢复合型 N-聚糖，而不是高甘露糖 N-聚糖。
A new mannose-recognizing lectin (MOL) was purified on an asialofetuin-column from fruiting bodies ofMarasmius oreadesgrown in Japan. The lectin (MOA) from the fruiting bodies of the same fungi is well known to be a ribosome-inactivating type lectin that recognizes blood-group B sugar. However, in our preliminary investigation, MOA was not found in Japanese fruiting bodies ofM. oreades, and instead, MOL was isolated. Gel filtration showed MOL is a homodimer noncovalently associated with two subunits of 13 kDa. The N-terminal sequence of MOL was blocked. The sequence of MOL was determined by cloning from cDNA and by protein sequencing of enzyme-digested peptides. The sequence shows mannose-binding motifs of bulb-type mannose-binding lectins from plants, and similarity to the sequences. Analyses of sugar-binding specificity by hemagglutination inhibition revealed the preference of MOL toward mannose and thyroglobulin, but asialofetuin was the strongest inhibitor of glycoproteins tested. Furthermore, glycan-array analysis showed that the specificity pattern of MOL was different from those of typical mannose-specific lectins. MOL preferred complex–type N-glycans rather than high-mannose N-glycans.