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Activation of Biofunctional Molecules Induced by Binding of Glycomolecules

糖分子结合诱导的生物功能分子的激活

基本信息

批准号：
08455435
负责人：
URYU Toshiyuki
金额：
$ 4.35万
依托单位：
UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

First, the reaction which binds oligosaccharides to peptides in mild conditions was examined. Malto-oligosaccharides having 2-7 glucose residues were found to the amino group of polylysine or polyarginine as protein model compounds by use of sodium cyanoborohydrate as a reducing agent at 37ﾟC for several hours. Since in NMR spectra of the reaction products, absorptions due to epsilon-carbon of poiyilsine side chains shifted from the original position at 40.9ppm to 50.2ppm, the ratio of each peak intensity was determined. Reaction conditions affording high degrees of substitution of 85-90% were found by using 10 equivalents of oligosaccharides such as maltose and maltopentaose.Next, bindings of oligosaccharides to natural enzymes such as aldolase, lactate dehydrogenase, 6-phosphogluconic dehydrogenase, cholinesterase, and phosphatase slkaline were performed. All sugar bound-enzymes except for aldolase (9%) exhibited58-90% of their original activities. In the case of maltose or maltoheptaose bound-phosphatase alkaline, it was found that, the enzyme activity after 10-20 days was much larger than that of the original phosphatase slkaline.In addition, to examine mechanisms of interactions of poilsaccharides with proteins, various mixtures of suifated polysaccharides such as curdlan sulfate, heparin, and chondroitin sulfate with polypeptide like polylysine, polyarginine, and their copolymers were measured by NMR.

首先，检查在温和条件下将寡糖结合至肽的反应。用氰基硼氢化钠作还原剂，在37 ℃下还原数小时，在作为蛋白质模型化合物的聚赖氨酸或聚精氨酸的氨基上发现了具有2-7个葡萄糖残基的麦芽低聚糖。由于在反应产物的NMR谱中，由于聚赖氨酸侧链的ε-碳的吸收从40.9ppm的原始位置移动到50.2ppm，因此确定了每个峰强度的比率。通过使用10当量的低聚糖如麦芽糖和麦芽五糖，发现了提供85-90%的高取代度的反应条件。接下来，进行了低聚糖与天然酶如醛缩酶、乳酸脱氢酶、6-磷酸葡萄糖酸脱氢酶、胆碱酯酶和磷酸酶slkaline的结合。除醛缩酶（9%）外，其它糖结合酶的活性均恢复到原来的58 -90%。此外，为了研究多糖与蛋白质相互作用的机制，将硫酸化多糖如硫酸凝胶多糖、肝素和硫酸软骨素与多肽如聚赖氨酸、聚精氨酸、聚赖氨酸和聚精氨酸的混合物进行了研究。并通过NMR测量它们的共聚物。