Protein-carbohydrate interactions : recognition and neural function of the mollusc (Aplysia) lectin.

因为我们找到了一个新的家庭鞘糖脂、phosphonoglycosphingolipids海兔kurodai,这些糖脂的结构和本地化进行了研究。针对FGL-IIb的抗血清仅对海兔的神经束染色，FGL-IIb特异性地集中在海兔的神经纤维中。因此，我们认为FGL-IIb可能识别细胞外基质上的某些成分。随访2年，结果如下：1994年，应用胆酸钠盐、Triton X-100和辛基葡萄糖苷三种洗涤剂对海兔神经组织进行溶解。本实验以辛基葡萄糖苷为最佳增溶剂，考察了上述洗涤剂对海芋神经组织中蛋白质的溶解度和糖脂的回收率。将经100 mM辛基葡萄糖苷处理的海兔神经组织溶解后的材料涂于抗fgl - iib抗体柱上，用50 mM辛基葡萄糖苷洗涤，0.1% SDS洗脱。通过SDS- page银染色分析，0.1% SDS洗脱层的45 kDa条带染色比正常IgG柱洗脱层更强烈。1995年，用氯仿/甲醇（2:1）从0.1% SDS洗脱液中提取的糖脂部分，仅用抗fgl - iib抗血清进行斑点免疫染色，未反应出其他糖脂的抗血清。另外0.1% SDS洗脱部分的SDS- page考马斯蓝染色为45 kDa带阳性。因此，我们报道了在细胞外基质上Ca^<2+>存在的情况下，特异性定位于海雀神经束的FGL-IIb与45 kDa蛋白形成复合物。45kda蛋白一级序列的研究正在进行中。

Since we found a new family glycosphingolipids, phosphonoglycosphingolipids, in Aplysia kurodai, the structures and the localizations of these glycolipids have been investigated. Antiserum against FGL-IIb, which is specifically concentrated in the nerve fibers of Aplysia, stained only nerve bundles of Aplysia. So, it was considered that FGL-IIb may recognize the some components on the extracellular matrix. Following results for 2 years were obtained.1. In 1994, Aplysia nervous tissues were solubilized by following three detergents, cholic acid sodium salt, Triton X-100 and octylglucoside. When the solubility of proteins and the recovery of glycolipids of Aplysia nervous tissues were investigated using above detergents, octylglucoside was best solubilizer in this experiment. Solubilized material of Aplysia nervous tissues treated with 100 mM octylglucoside was applied to anti-FGL-IIb antibody column and the column was washed with 50 mM octylgucoside and then eluted with 0.1% SDS solution. Silver staining by SDS-PAGE analysis of the 0.1% SDS eluate stained 45 kDa band more strongly than normal IgG column eluate.2. In 1995, glycolipid fraction that was extracted with chloroform/methanol (2 : 1) from the 0.1% SDS eluate fraction was positively dot immunostained only by anti-FGL-IIb antiserum, but antisera against other glycolipids were not reacted. Coomassie Blue staining by SDS-PAGE of the another 0.1% SDS eluate fraction stained positively 45 kDa band.Therefore, we reported that FGL-IIb which specifically localized in Aplysia nerve bundles formed the complex with 45 kDa protein in the presence of Ca^<2+> on the extracellular matrix. Study on the primary sequence of the 45 kDa protein is in progress.