Proteomic analysis of the pathogenic domain related to aggregate formation

我们对与聚集体形成有关的结构域进行了研究，特别是对筏和聚谷氨酰胺聚集体本身进行了研究。主要研究结果如下：1.建立了蛋白质特异性筏式分离方法; 2.鉴定了P62为一种蛋白质，其含量随着多聚谷氨酰胺聚集体的形成而增加; 3.建立了不使用强去污剂的多聚谷氨酰胺聚集体分离方法，并进行了聚集体相关蛋白（AIP）的质谱分析。我们在AIP中发现了几种已知和未知的蛋白质，其中包括泛素结合蛋白ubiquilin。4.我们发现钠通道β亚基4在亨廷顿病脑中是一种减少蛋白。5. β亚基被鉴定为BACE 1和γ分泌酶的新底物。6.我们制备了亨廷顿蛋白外显子1与EGFP融合的转基因小鼠，以观察聚集体。

We studied on the domain, which related to aggregate formation, especially on raft and polyglutamine aggregate itself. The followings are the results which we obtained in this projects.1.The protein specific raft isolation method was established.2.P62 was identified as a protein, which increased as the polyglutamine aggregates were formed.3.We established isolation method of polyglutamine aggregate without using strong detergent and mass spectrometry analysis was performed to identify aggregate associated proteins(AIP). We found several known and unknown proteins in AIP including ubiquilin, one of the ubiquitin binding proteins.4.We found sodium channel beta subunit 4 as a decrease protein in Huntington disease brain. The beta subunits exist in the raft fraction in the brain.5.Beta subunits are identified as new substrates for both BACE1 and gamma secretase.6.We made transgenic mouse of huntingtin exon1 fused with EGFP to visualize the aggregates.