Proteomic analysis of the pathogenic domain related to aggregate formation

我们研究了与聚集体形成有关的结构域，特别是RAFT和聚谷氨酰胺聚集体本身。1.建立了蛋白质特异性RAFT分离方法。2.鉴定了P62是一种蛋白质，随着聚谷氨酰胺聚集体的形成而增加。3.建立了不使用强洗涤剂的聚谷氨酰胺聚集体的分离方法，并对聚集态相关蛋白(AIP)进行了鉴定。我们在AIP中发现了几个已知和未知的蛋白，包括泛素结合蛋白之一泛素蛋白。4.我们发现钠通道β亚基4是亨廷顿病脑内的一个减少蛋白。5.β亚基被鉴定为BACE1和伽马分泌酶的新底物。6.我们制作了亨廷顿蛋白外显子1与绿色荧光蛋白融合的转基因小鼠，以显示聚集体。

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.