摘要
Acidic oligosaccharide sugar chain (AOSC), a D-mannuronic acid oligosaccharide, derived from brownalgaepolysaccharide, has been completed Phase I clinical trial in China as an anti-Alzheimer's Disease (AD) drug candidate. The identification of AOSC-binding protein(s) in neurons is very important for understanding its action mechanism. To determine the binding protein(s) of AOSC in neurons mediating its anti-AD activities, confocal microscopy, affinity chromatography, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis were used. Confocal microscopy analysis shows that AOSC binds to SH-SY5Y cells in concentration-, time-, and temperature-dependent fashions. The AOSC binding proteins were purified by affinity chromatography and identified by LC-MS/MS analysis. The results showed that there are 349 proteins binding AOSC, including clathrin, adaptor protein-2 (AP-2) and amyloid precursor protein (APP). These results suggest that the binding/entrance of AOSC to neurons is probably responsible for anti-AD activities.
酸的 oligosaccharide 糖链(AOSC ) , D-mannuronic 酸 oligosaccharide,源于棕色的水藻多糖,被完成了阶段我在是的中国的临床的试用 anti-Alzheimer 的疾病(广告) 药候选人。在神经原的 AOSC 有约束力的蛋白质的鉴定为理解它的行动机制是很重要的。为了在调停的神经原决定 AOSC 的有约束力的蛋白质,它的反广告活动,共焦的显微镜学,亲密关系层析,和液体层析双人脚踏车集中 spectrometry (LC-MS/MS ) 分析被使用。AOSC 在集中绑在 SH-SY5Y 房间的共焦的显微镜学分析表演 -- ,时间 -- ,并且温度依赖者时尚。AOSC 有约束力的蛋白质被亲密关系层析净化并且由 LC-MS/MS 分析识别了。结果证明有 349 蛋白质绑定 AOSC,包括 clathrin,适配器 protein-2 (AP-2 ) 和淀粉的先锋蛋白质(应用软件) 。这些结果建议到神经原的 AOSC 的绑定 / 入口为反广告活动可能负责。
基金
Supported by National Basic Research Program of China (973 Program) (No.2003CB716400)
Natural Science Foundation of China for Distingutshed Young Scholars (No.30725046)
Natural Science Foundation of China (No.30472029,No.30772612)
Chinese High technology Research and Development Program of China(863 Program) (No.2006AA090501)