Biological Activity of novel glycosaminoglycan and interaction with binding proteins.

新型糖胺聚糖的生物活性及其与结合蛋白的相互作用。

基本信息

批准号：
09672185
负责人：
TOIDA Toshihiko
金额：
$ 1.98万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672185/
关键词：
Acharan sulfate oligosaccharides acticoagulant activity interaction glycosaminoglycan giant snail NMR spectroscopy affinity

项目摘要

We have very recently found a new glycosaminoglycan from African Giant Snail, Achatina fulica, and have characterized that this new glycosaminoglycan is consisting of homogeneous disaccharide (*4GlcNpAcalpha1*4IdoAp2Salpha) repeating unit. This polysaccharide, named acharan sulfate, has a unique sequence belonging in heparan sulfate family and shows angeogenesis activity.This study examined the hypothesis that contain oligosaccharide sequences in acharan sulfate have high affinity for consensus peptide sequences in proteins present in the extracellular matrix. A variety of different size of oligosaccharides in the range from disaccharide to tetradecasaccharide of acharan sulfate were prepared by partial enzymatic digestion. These oligosaccharides were used to isolate high affinity proteins, which were examined for composition and sequence. Oligosaccharide-protein interactions were examined for : 1. binding affinity (Kd) ; 2. binding specificity ; 3. Binding thermodynamics ; 4. binding … More orientation ; and 5. binding site structural requirements! Peptides will also be synthesized to contain variations of the consensus sequence and modified oligosaccharides such as N-de-acetylated and re-N-sulfonated acharan sulfate, were synthesized to help establish the structure activity relationship for oligosaccharide-peptide interaction. Native protein will be compared with synthetic peptide to confirm specificity of binding. Vitronectin and recombinant acidic fibroblast growth factor are the proteins to be studied because of the important biological consequences of their binding to acharan sulfate/heparin/heparan sulfate. Ultimately, these studies should lead to custom designed peptides that have increased oligosaccharide binding affinity to enhance or attenuate the activities associated with oligosaccharide-protein interaction. These oligosaccharides and peptides represent the starting point for new drug design focused on peptido-mimetics and sulfated oligosaccharide-mimetics. Such new therapeutic agents might be used to regulate a variety of biological processes including angiogenesis, metastasis, atherogenesis, immune responses, HIV infection, and thrombosis. Less

我们最近从非洲巨型蜗牛Achatina Fulica找到了一种新的糖胺聚糖，并且已经表明，这种新的糖胺聚糖由均匀的二糖组成（*4Glcnpacalpha1*4glcnpacalpha1*4idoap2salpha）重复单元。这种称为Acharan硫酸盐的多糖具有独特的序列，属于硫酸乙酰肝素家族，并显示出序列化活性。本研究研究了一个假说，即在硫酸achar硫酸achar素硫酸盐中含有寡糖序列对蛋白质中蛋白质肽序列的高亲和力具有高亲和力。从二糖到硫酸阿查兰的四氯，在硫酸盐的四碳中，各种不同大小的寡糖都是通过部分酶促消化制备的。这些寡糖用于分离高亲和力蛋白，该蛋白被检查是否有组成和序列。检查了：1。结合亲和力（KD）的寡糖蛋白相互作用； 2。结合特异性； 3。结合热力学； 4。绑定…更多方向；和5。绑定站点结构要求！还将合成肽含有共识序列的变化，并合成诸如N-脱乙酰化和重新磺化的硫酸盐的寡糖变化，并合成以建立寡糖肽相互作用的结构活性关系。将天然蛋白与合成肽进行比较，以确认结合的特异性。玻璃原蛋白和重组酸性成纤维细胞生长因子是要研究的蛋白质，因为它们与硫酸阿查兰/肝素/肝素硫酸盐的结合产生了重要的生物学后果。最终，这些研究应导致定制设计的肽，这些肽增加了寡糖结合亲和力，以增强或减轻与寡糖 - 蛋白质相互作用相关的活性。这些寡糖和Petides代表了新药设计的起点，该设计集中在辣椒摄影和硫化寡糖摄影上。这种新的治疗剂可用于调节各种生物学过程，包括血管生成，转移，动脉粥样硬化，免疫反应，HIV感染和血栓形成。较少的