SARS冠状病毒主蛋白酶和S蛋白的结构与功能研究及其抑制剂的设计

发布时间：2018-01-14 15:29

本文关键词：SARS冠状病毒主蛋白酶和S蛋白的结构与功能研究及其抑制剂的设计　出处：《浙江大学》2005年博士论文　论文类型：学位论文

【摘要】：传染性非典型肺炎(Severe Acute Respiratory Syndrome,SARS)自爆发以来给全世界尤其是中国带来了重大的人身伤害和经济破坏,本课题组及时发挥本组在药物设计方面的多年经验积累,在抗SARS药物设计以及免疫方面开展了积极而有成效的研究,其主要内容由以下几部分工作组成: 1、SARS病毒基因组主要编码的蛋白质有RNA聚合酶蛋白(聚合酶1a,1b),S蛋白(spike protein),E蛋白(small membrane protein),N蛋白(nucleocapsid protein),主蛋白酶(3C-like proteinase)等。其中,主蛋白酶以其在SARS病毒复制的过程中所起的重要作用而成为研究的热点。我们利用先进的生物信息学分析软件从已知结构的蛋白质数据库(PDB)中搜寻到猪冠状病毒(TGEV)和人感冒冠状病毒(HCoV)主蛋白酶与SARS病毒主蛋白酶序列具有很高相似性。基于此基础,以TGEV的主蛋白酶三维晶体结构为模板来同源模建了SARS冠状病毒的主蛋白酶三维结构,分析了其结构和活性位点特征。 2、由于实验测得TGEV和HCoV主蛋白酶在溶液中均有二聚体的形式存在,并且可能在发挥蛋白酶生物活性方面具有重要作用,因此,我们初步推测SARS病毒主蛋白酶在体内也具有二聚体的存在。参考TGEV和HCoV主蛋白酶二聚体形式,首先搭建了SARS病毒主蛋白酶的二聚体。研究发现,三种蛋白二聚体界面处的静电分布具有明显的互补性,三种蛋白酶二聚体单体之间具有相同的静电和疏水相互作用能。表明SARS病毒主蛋白酶在溶液中也具有二聚体的存在,这被后期的实验所证实。 3、近期有实验证实SARS病毒主蛋白酶二聚体为该蛋白酶主要生物形式,而单体不具有活性。进一步测得了该蛋白酶的二聚体晶体结构,但是其单体的结构不能测得,这也就使单体失活的原因不能知晓。因此,本文采用分子动力学模拟方法对该蛋白酶的二聚体和单体形式分别作了1.8ns的分动力学研究,来考察蛋白单体和二聚体形式构象的变化。通过该动力学模拟发现,SARS病毒主蛋白酶二聚体活性口袋具有如下三个特征:1)活性位点His41和Cys145间形成离子
[Abstract]:Severe Acute Respiratory Syndrome. Since the outbreak of SARSs to the world, especially to China has brought major personal injury and economic damage, this group in a timely manner in drug design in many years of experience accumulated. Active and effective studies have been carried out on the design of anti- SARS drugs and immunization, the main contents of which are composed of the following parts: 1 the main protein encoded in the genome of SARS-CoV is RNA polymerase (1a ~ 1b) spike protein. E protein, small membrane protein, N protein, nucleocapsid protein. Main protease 3C-like proteinase. The main protease has become a hot topic for its important role in the replication of SARS virus. We use advanced bioinformatics analysis software to analyze the protein database with known structure. Based on this basis, the sequence of major protease of porcine coronavirus (TGEV) and human common cold coronavirus (HCoV) was similar to that of SARS virus. The three-dimensional structure of SARS coronavirus was constructed by using the three-dimensional crystal structure of main protease of TGEV as template, and the structure and active site characteristics of SARS coronavirus were analyzed. 2. It was found that both TGEV and HCoV have dimer in solution and may play an important role in the biological activity of protease. We preliminarily speculate that the main protease of SARS virus also has dimer in vivo. We refer to the TGEV and HCoV main protease dimer forms. First, the dimer of the main protease of SARS virus was constructed. It was found that the electrostatic distribution at the interface of the three protein dimers was obviously complementary. The three protease dimer monomers have the same electrostatic and hydrophobic interaction energy, which indicates that the main protease of SARS virus also has dimer in solution, which was confirmed by later experiments. 3. Recent experiments have confirmed that the main protease dimer of SARS virus is the main biological form of the protease, but the monomer has no activity. The dimer crystal structure of the protease was further measured. However, the structure of the monomer can not be measured, which makes the cause of the monomer inactivation unknown. In this paper, the dimer and monomer form of the protease were studied by molecular dynamics simulation method. To investigate the conformation changes of monomers and dimers. The active pockets of the main protease dimer of SARS virus have the following three characteristics: 1) the ions formed between His41 and Cys145
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R373

【相似文献】