β-淀粉样蛋白与膜脂作用后的交联研究

发布时间：2019-02-09 16:05

【摘要】：β淀粉样蛋白(Aβ)与细胞膜相互作用形成寡聚体存在于膜脂内,聚丙烯酰胺凝胶电泳常用于分析这些膜内寡聚体的结构和状态,但十二烷基硫酸钠(SDS)使得寡聚体原有的结构极易被破坏,使用合适的交联剂来稳定蛋白在膜上的状态,从而获取寡聚体本身的结构.用双琥珀酰亚胺辛二酸酯磺酸钠盐(BS3)对Aβ42进行交联,将BS3与戊二醛交联进行比较分析,通过电泳、脂质体技术,并采用电镜照片对照分析.实验结果显示:1)BS3交联后的样品能保持蛋白原先的状态;2)BS3不会导致蛋白过度交联,很适合作为分析膜脂与β淀粉样蛋白相互作用的交联剂.因此,报道了BS3分析β淀粉样蛋白这种易聚集蛋白在插膜后的结构,且BS3可以替代戊二醛成为更有效的交联剂,这将为β淀粉样蛋白及其他易聚集蛋白插膜提供很好的研究方法.
[Abstract]:尾 -amyloid protein (A 尾) interacts with cell membrane to form oligomers in membrane lipids. Polyacrylamide gel electrophoresis is often used to analyze the structure and state of these oligomers. However, sodium dodecyl sulfate (SDS) can easily destroy the original structure of the oligomer, so the structure of the oligomer can be obtained by using suitable crosslinking agent to stabilize the state of the protein on the membrane. The A 尾 42 was crosslinked by sodium succinimide octanedioate sulfonate (BS3). The crosslinking of BS3 and glutaraldehyde was compared and analyzed by electrophoresis, liposome technique and electron microscopy. The results showed that: 1) BS3 cross-linked samples could maintain the original protein status, 2) BS3 would not lead to excessive cross-linking of proteins, and was suitable for the analysis of the interaction between membrane lipids and 尾 -amyloid proteins. Therefore, BS3 was used to analyze the structure of 尾 -amyloid protein after intercalation, and BS3 could replace glutaraldehyde as a more effective crosslinking agent. This will provide a good method for 尾-amyloid and other easy-aggregation protein intercalation.
【作者单位】：西藏民族大学基础医学院/高原环境与疾病相关基因研究实验室;
【基金】：西藏自然科学基金项目(12KJZRZMY02)
【分类号】：R749.16

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