Expansion of structure biology studies on human defense and response proteins for drug design based on three-dimensional-structures

扩展人类防御和反应蛋白的结构生物学研究，用于基于三维结构的药物设计

基本信息

批准号：
14370727
负责人：
SATOW Yoshinori
金额：
$ 8.77万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

For drug design based on three-dimensional structures of defense and response proteins such as human receptors and disease-related enzymes, structural biology studies utilizing X-ray crystallography are carried out on these proteins. The studies have further expanded the methods for structure study and their application.Bacterial Lipopolysaccharide is recognized by human MD-2 proteins that associated with human Toll-Like receptor 4 (TLR4). The TLR4 complex with MD-2 transmits bacterial infection signals to cell cytosol, triggers defense response to the infection, and causes endotoxin shocks known as septicemia. Human and mouse MD-2 proteins are expressed in yeast and then highly purified. Crystals are obtained for human MD-2. Attempts to express human TLR4 are also carried out. Human heat-shock protein HSP40 and β2 trans-membrane receptor are also expressed, purified, and subjected to crystallization attempts.Human β-galactosidasese and α-N-acetylgalactosaminidase are lysosomal enzymes … More hydrolyzes galactoside linkages in various glycolipids, and its genetic mutations decrease enzymatic activities which result in deficiencies of glycolipid metabolism and hence accumulate substrates in organs, causing lysosomal diseases. These enzymes are expressed and subjected to crystallographic studies. The structure of human tissue factor protein in complex with humanized antibody Fab is determined for drug design of blood-coagulation inhibitorsYeast homing endonuclease derived from VMA1 gene is structurally studied for full elucidation of its binding to double-stranded substrate DNAs. To elucidate its precise reaction mechanisms of protein splicing in this endonuclease, a series of crystal structural studies has been carried out with the use of splicing-inactive and slowly spliceable precursors. Pathogenic protein SEp22 from Salmonella enteritidis is expressed, purified, and crystallized. The structure of SEp22 determined for two crystal forms has revealed 12-subunitit oligomer in 23 symmetry and iron ion binding for protection of its DNA. Cassette recombinases of Staphylococcus aureus (SA) are expressed and purified for understanding of methicillin-resistant SA diseases. Less

对于基于防御和反应蛋白的三维结构（例如人体受体和疾病相关酶）的药物设计，对这些蛋白质进行了利用X射线晶体学的结构生物学研究。这些研究进一步扩大了结构研究及其应用的方法。细菌脂多糖是由与人Toll样受体4（TLR4）相关的人类MD-2蛋白认识到的。带有MD-2的TLR4复合物将细菌感染信号传播到细胞胞质，触发对感染的防御反应，并引起内毒素休克，称为败血病。人和小鼠MD-2蛋白在酵母中表达，然后高度纯化。对于人类MD-2获得了晶体。也进行了表达人类TLR4的尝试。人类热休克蛋白HSP40和β2跨膜受体也被表达，纯化和进行结晶尝试。人类β-半乳糖苷酶和α-N-N-乙酰乳糖苷酶酶是溶酶体是溶酶体的……更多的水解酶在各种水解酶上均在各种孔酰基中构成突变，并构成了概括的概念突变，该突变构成了概括的概念突变。糖脂代谢以及器官中的丙烯酸底物的，引起溶酶体疾病。这些酶表示并进行晶体学研究。与人源化抗体Fab复合物中的人体组织因子蛋白的结构是用于血液凝结的药物设计的抑制剂抑制作用的归巢核酸内切酶，该核酸酶源自VMA1基因是结构研究的，用于完全阐明其与双链底物DNA的结合。为了阐明其在此核酸酶中蛋白质剪接的精确反应机制，使用剪接 - 无效和可缓慢剪接的前体进行了一系列晶体结构研究。从肠道沙门氏菌中的致病蛋白SEP22表达，纯化和结晶。针对两种晶体形式确定的SEP22的结构揭示了23个对称性和铁离子结合以保护其DNA的12个亚基低聚物。葡萄球菌（SA）的盒式重组酶表示并纯化，以理解耐甲氧西林的SA疾病。较少的