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

蛋白质-碳水化合物相互作用：软体动物（海兔）凝集素的识别和神经功能。

基本信息

批准号：
06680755
负责人：
ABE Sachiko
金额：
$ 1.54万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

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.

自从我们在黑大杏中发现了一个新的鞘糖脂家族--膦酰鞘糖脂以来，人们对这些糖脂的结构和定位进行了研究。抗FGL-IIb的抗血清，这是专门集中在神经纤维的Aaplasia，染色神经束的Aaplasia。因此，认为FGL-IIb可能识别细胞外基质上的某些成分。2年来取得了如下结果. 1994年用胆酸钠盐、Triton X-100和辛基葡萄糖苷三种去污剂溶解了阿氏藻神经组织。在本实验中，辛基葡萄糖苷是最佳的增溶剂。将用100 mM辛基葡糖苷处理的Aaplasia神经组织的溶解材料施加到抗FGL-IIb抗体柱上，用50 mM辛基葡糖苷洗涤柱，然后用0.1%SDS溶液洗脱。银染经SDS-PAGE分析的0.1%SDS PAGE染色的45 kDa条带比正常IgG柱洗脱液更强. 1995年，从0.1%SDS-PAGE级分中用氯仿/甲醇（2：1）提取的糖脂级分仅被抗FGL-IIb抗血清阳性斑点免疫染色，而其它糖脂抗血清不反应。SDS-PAGE结果表明，FGL-Ⅱ b蛋白在细胞外基质中与45 kDa蛋白形成复合物，该复合物特异性定位于神经束中，并与细胞外基质中的Ca^<2+>结合。对45 kDa蛋白的一级序列的研究正在进行中。