Protein Structure

蛋白质结构

基本信息

批准号：
7592679
负责人：
alexander wlodawer
金额：
$ 165.62万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7592679
关键词：
ATP-Dependent Proteases Acquired Immunodeficiency Syndrome Antibodies Antiviral Agents Archaeoglobus fulgidus Area Aspartic Endopeptidases Binding Biogenesis Biological C-terminal Carbohydrates Complex Crystallography Cytokine Receptors Disaccharides Drug resistance Endopeptidases Enzymes Escherichia coli Family Feline Immunodeficiency Virus HIV Infections HIV Protease Interleukin-10 Interleukin-15 Investigation Laboratories Lectin Malignant Neoplasms Mannose Monosaccharides N-terminal Numbers Organism Peptide Hydrolases Pharmaceutical Preparations Phosphorylation Phosphotransferases Process Property Protease Domain Protein-Serine-Threonine Kinases Proteins RNA, Ribosomal, 18S Research Resolution Ribosomes Role Serine Site Source Structure Structure-Activity Relationship Techniques Translation Initiation Tumor Suppression Tumor Suppressor Proteins Variant Work X ray diffraction analysis X-Ray Diffraction allergen Bla g 2 base cockroach allergen cytokine endopeptidase La inhibitor/antagonist interest interleukin 20 interleukin-19 interleukin-22 method development novel preclinical study prevent programs protein structure protein structure function receptor

项目摘要

In the past several years, our work has concentrated in four distinct areas. Crystallographic studies of proteases Crystallographic studies of proteases have been an important area of research of this Section since its establishment. We have been particularly active in the investigation of structure-function relationship in aspartic proteases, including clinically important retroviral enzymes. Our studies of HIV protease, although no longer a major target of active research, are still ongoing and concentrate on the investigation of drug-resistant variants and their complexes with inhibitors. We have investigated retroviral proteases from several other sources such as FIV, RSV, and HTLV, having recently solved the structure of the latter enzyme. Cockroach allergen Bla g 2 was shown to be an inactive aspartic protease and we solved its structure in a complex with a specific antibody. We have established an extensive program of investigating serine-carboxyl peptidases (sedolisins), a family that was first characterized based on crystal structures solved in this laboratory and that is found in many different organisms. We are also investigating a bacterial ATP-dependent protease Lon, finding that is proteolytic domain has a unique fold and thus establishes a new family of proteases with a Ser-Lys catalytic dyad. We have solved the structures of the proteolytic domain of A and B type Lon proteases, encoded by E. coli and Archaeoglobus fulgidus, as well as the N-terminal and alpha domains of E. coli Lon. Lectins with antiviral activity We have been involved in studies of several lectins with antiviral activities, some of them currently being in pre-clinical trials as potential drugs preventing HIV infection. We have solved the structure of griffithsin, as free protein and complexed with a number of mono- and disaccharides, explaining the structural basis for its tight binding to branched mannose-rich carbohydrates. We have also solved atomic-resolution structure of another lectin, scytovirin. Proteins involved in ribosome biogenesis and tumor suppression Two related serine protein kinases, Rio1 and Rio2, are involved in processing 20S pre-RNA to 18S ribosomal RNA. Their crystal structures, solved by our Section, established that they belong to a novel family of kinases with a truncated substrate-binding region, although they are capable of both self- and trans-phosphorylation. We have established the sites of autophosphorylation for both of them. We are currently investigating their catalytic properties and a potential biological role. We have also solved the structure of the C-terminal MA3 domain of Pdcd4, explaining how that tumor suppressor factor inhibits translation initiation. Cytokines and cytokine receptors Our Section has been investigating the crystal structures of several cytokines and has made progress in preparing their receptor complexes. We have purified and crystallized complexes of IL-10 with its specific receptor and are studying complexes of several other cytokines related to IL-10, such as IL-19, IL-20, and IL-22. We are also investigating structural features of the interactions of IL-15 with the receptor molecules.

在过去的几年中，我们的工作集中在四个不同的领域。蛋白酶的晶体学研究蛋白酶的晶体学研究一直是自建立以来本节的重要领域。我们在天冬氨酸蛋白酶（包括临床上重要的逆转录病毒酶）中对结构功能关系的研究特别活跃。我们对HIV蛋白酶的研究虽然不再是积极研究的主要靶标，但仍在进行中，并集中于对抗药性变体及其与抑制剂的复合物进行研究。我们已经研究了来自FIV，RSV和HTLV等其他几个来源的逆转录病毒蛋白酶，最近解决了后一种酶的结构。蟑螂过敏原Bla G 2被证明是一种非活性天冬氨酸蛋白酶，我们在具有特定抗体的复合物中求解了其结构。我们已经建立了一个广泛的计划，以研究丝氨酸 - 羧基肽酶（Sedolisins），这是一个基于在该实验室中解决的晶体结构而首先表征的，并且在许多不同的生物体中发现。我们还正在研究细菌依赖ATP的蛋白酶lon，发现蛋白水解结构域具有独特的折叠，因此建立了具有Ser-Lys催化二元组的新蛋白酶家族。我们已经解决了由大肠杆菌和fulgidus编码的A和B型LON蛋白酶的蛋白水解结构域的结构，以及大肠杆菌lon的N端和α域的结构。具有抗病毒活性的凝集素我们参与了几种具有抗病毒活性的凝集素的研究，其中一些目前正在临床前试验中，作为预防HIV感染的潜在药物。我们已经解决了griffithsin的结构，作为游离蛋白质，并与许多单糖和二糖构成复合，从而解释了其与富含甘露糖的富含甘露糖的碳水化合物紧密结合的结构基础。我们还解决了另一种凝集素Scytovirin的原子分辨率结构。参与核糖体生物发生和肿瘤抑制的蛋白质两种相关的丝氨酸蛋白激酶RIO1和RIO2参与将20S前RNA处理至18S核糖体RNA。他们的晶体结构由我们的部分解决，确定它们属于具有截短的底物结合区域的新型激酶家族，尽管它们既具有自我和反磷酸化。我们已经为两者建立了自磷酸化的位置。我们目前正在研究它们的催化特性和潜在的生物学作用。我们还解决了PDCD4的C末端MA3结构域的结构，并解释了该肿瘤抑制因子如何抑制翻译起始。细胞因子和细胞因子受体我们的截面一直在研究几种细胞因子的晶体结构，并在制备其受体复合物方面取得了进展。我们已经纯化和结晶的IL-10及其特定受体的复合物，正在研究与IL-10相关的其他几种细胞因子的复合物，例如IL-19，IL-20和IL-22。我们还正在研究IL-15与受体分子相互作用的结构特征。