兰州大学机构库 >生命科学学院
淀粉样前体蛋白转运机制的研究
Alternative TitleThe Study on Amyloid Precursor Protein's Trafficking Mechanism
吕玲玲
Thesis Advisor吉尚戎
2016-06-01
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword阿尔兹海默氏症 淀粉样前体蛋白 固有无序结构域 免疫荧光
Abstract阿尔茨海默氏病(Alzheimer’s disease, AD) 是一种对患者乃至患者家人都有极大负面影响力的神经退行性疾病。越来越多的生物化学和遗传学证据显示淀粉样前体蛋白(APP , Amyloid Precursor Protein) 在AD的发病机制中奠定了举足轻重的作用。APP是I型跨膜蛋白质,它类似于一种细胞表面受体,并包含一个大的胞外N端结构域和一个短的胞质尾。虽然APP的研究集中于其C端,但大量的文献表明APP的N端能够调节其自身的转运和加工。而其含有的固有无序区更增加了其调节自身行为的困难性。近年来对固有无序结构域功能的研究逐渐增多,其不仅可以影响蛋白的半衰期,而且可以适时地改变蛋白结构,调节蛋白和其他蛋白的相互作用等。利用瞬时表达等方法,我们发现APP无序区ACD的缺少,可以使得APP被酶加工后,分泌型的APP减少。不同抗体检测、胞外不同长度SAPP检测、胞内SAPP检测均证明该现象的稳定性。为了进一步探究胞外分泌减少的原因,我们使用免疫荧光共定位的手段,发现在内质网中野生型和突变体无差异,在高尔基体处缺少无序区的APP与野生型的APP差异极显著,删除ACD结构域的APP蛋白在trans-高尔基体处并未出现如野生型APP一般的定位升高。利用APP蛋白的N端抗体与早期内吞体的共定位发现删除ACD结构域并不影响APP及SAPP在内吞体中分布的总量。粗提脂筏实验从蛋白定位的微环境阐述了删除ACD结构域引起了APP蛋白在细胞表面定位的差异。至此我们可以得出APP胞外域可以通过影响APP蛋白在trans-高尔基体中的分布、蛋白在细胞表面的定位等途径来调节APP的转运。
Other AbstractAlzheimer's diseases (AD) which acts as a neurodegenerative disease, has great negative influence to patients and patients' family. More and more biochemical and genetic evidences show that amyloid precursor protein (APP) has a very important role in the pathogenesis of AD. APP is a type I transmembrane protein which is similar to a cell surface receptor, with a large extracellular N-terminal structural domain and a short cytoplasmic tail. Although the study of the APP focuses on the C-terminal, a large number of evidences show that the N-terminal of the APP is able to regulate the trafficking and processing of its own. Inherent disordered area within the N-terminal also increases the possibility and mystery of the regulating. In recent years, the research on the inherent disorder protein(IDP) show that it can affect the protein half-life, timely protein structure changing and protein-protein interactions regulation. By using the methods of transient expression, we found that the lack of IDP could reduce the production of SAPP. In order to further explore the underlying mechanism of this phenomenon, by using intensity correlation quotient (ICQ) showed the colocalization with golgi apparatus was significantly different between the wild type APP and the disorder domain deletion APP, whereas in endoplasmic reticulum there was no difference, and the disorder domain deletion APP quantity did not elevate at trans-golgi apparatus like the wild type. We also found the deletion of ACD domain was not going to effect the quantity of APP and SAPP in endosome with N-terminal antibody colocalization. The crude lipid raft extraction experiments indicated that the deletion of ACD domain could cause distribution changing of APP at the surface of the cell from the perspective of the micro-environment of protein location. Until now, we can draw the conclusion that the extracellular domain of APP could regulate the trafficking and processing of APP by APP’s distribution in trans-golgi apparatus and location at the cell surface.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221535
Collection生命科学学院
Recommended Citation
GB/T 7714
吕玲玲. 淀粉样前体蛋白转运机制的研究[D]. 兰州. 兰州大学,2016.
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