cypj蛋白晶体结构解析及其抑制剂的筛选和鉴定-医学遗传学专业毕业论文.docx

博士后出站报告 CyPJ蛋白晶体结构解析及】E抑制剂的筛选和鉴定
殖,、±。
关键词:亲和素J、肽酰脯氨酰顺反异构酶、三维结构、小分子数据库、分子相
互作用分析仪、半抑制浓度
复旦大学 生物学博士后流动站 2
博上后出站报告毋P】蛋白晶体结构解析发其抑制剂的筛选和鉴定
Abstract
prise a highly abundant family of proteins that are expressed in anism studied to have been shown to act as peptidyl-prolyl c/s-trans isomerase whose activity can be inhibited by novel member cyclophilin J in this family was cloned,expressed and purified,the structure of CyPJ protein。the inhibitors of GyPJ protein,and the biological function of those inhibitors
was obtained and analyzed.
(1)The three-dimensional structure of has been determined by molecular replacement using the CyPA structure as the search model and has been refined at CyPJ molecule contains four helices and one B—posed of eight antiparallel 8- over secondary and tertiary structures of CyPJ are similar to those of CyPA,but CyPJ contains an additional disulfide bridge and four segments with conformations that are strikingly different from those of and Gin52 of CyPJ
are expected to be the active sites based on sequence alignment with CyPJ
structure shows a conserved water molecule wat226 close to His43 and GIn52.
(2)In Chapters On which virtual screening was carried OUt to discover novel inhibitors from the available small molecule the virtual screening,bined strategy of pharmacophore—based database search(PBDS)with docking。based database search was using PBDS a focus database was constructed by filtering out most of the possibly pounds from the large database;and by using docking method candidates that are possible binders of CyPJ were pick out from the small molecule database. BIACORE assay revealed that,of the 63 purchased
compounds 1 8 are active in binding CyPJ.
(3)The PPlase activity of CyPJ and CyPJ protein—pounds interaction were determined by o—chymotrypsin-coupled enzymic result