唐氏综合症决定域蛋白1泛素化分析

利用Fm oc固相多肽合成的方法合成DSCR1羧基端一个多肽片段(55-70AA),经HPLC纯化后偶联到匙孔槭血蓝蛋白,免疫新西兰雄兔后采血检测、纯化、经W estern b lotting、免疫沉淀证实得到的抗体为抗DSCR1的特异抗体。该抗体即能检测人源DSCR1蛋白,又能检测小鼠的DSCR1蛋白。运用获得的DSCR1多克隆抗体进行功能研究,发现DSCR1广泛存在泛素化,参与泛素化-蛋白酶体途径。

DSCR1 may be Participated in Ubiquitin-proteosome Pathway

Human DSCR1 gene is located in Down syndrome critical region on chromosome 21.The DSCR1 expressed extensively in body such as breast,uterus,skin,fetus,lung,liver and pancreas that distributed with different degree.Now,there is growing evidence that DSCR1 is involved in not only Down syndrome but also Alzheimer's disease.However,we know little about its detailed roles in these diseases so far.In this work,to synthesize the differential immunogenic peptide,then the purified peptide was coupled to the hemocyanin from Keyhole Limpets(KLH).The peptide-KLH solution was mixed with freund's complete or incomplete adjuvant and immunized rabbit to prepare specific polyclonal antibody against the N-terminus 5570 amino acids segment of DSCR1 large isotype.Western blotting analysis and immunoprecipitation suggested that the polyclonal antibody recognize the DSCR1 protein in murine brain and heart.DSCR1 protein was ubiquitously modified by the experiment of dealing Hela cells with MG132.So the DSCR1 may be involved in "ubiquitin-proteosome" pathway.Then we developed co-immunoprecipitation combined DSCR1 polyclonal antibody with ubiquitin polyclonal antibody to detect murine brain tissue.The result suggested that DSCR1 protein in mouse brain had ubiquitous modification indeed.