Protein Structure

蛋白质结构

基本信息

批准号：
7965279
负责人：
alexander wlodawer
金额：
$ 177.47万
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/7965279
关键词：
ATP-Dependent Proteases Acquired Immunodeficiency Syndrome Antibodies Antiviral Agents Area Aspartic Endopeptidases Binding Carbohydrates Complex Crystallography Cytokine Receptors Data Databases Deposition Development Disaccharides Drug resistance Enzymes Family Feline Immunodeficiency Virus HIV Infections HIV Protease Human T-lymphotropic virus 1 Interferons Interleukin-10 Investigation Laboratories Lectin Malignant Neoplasms Mannose Methodology Monosaccharides Oligosaccharides Organism Peptide Hydrolases Pharmaceutical Preparations Phase Property Protease Domain Proteins Research Resolution Serine Source Structure Structure-Activity Relationship Techniques Variant Work X ray diffraction analysis X-Ray Diffraction allergen Bla g 2 base cockroach allergen cytokine drug development inhibitor/antagonist interest interleukin 20 interleukin-19 interleukin-22 leukemia method development preclinical study prevent programs protein complex 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-1. A number of inhibitor complexes of the latter enzyme have been analyzed, with the aim of assisting in the development of drugs against HTLV-caused leukemia. Cockroach allergen Bla g 2 was shown to be an inactive aspartic protease and we solved the structures of two complexes with different specific antibodies. 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. 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 reengineered griffithsin into a monomeric form and solved its structure with a complex oligosaccharide, elucidating the basis of its antiviral properties. We have also solved atomic-resolution structure of another lectin, scytovirin. 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 have solved the structure of lambda interferon complexed with its receptor. Development of crystallographic methodology We have been investigating the problems related to phasing of diffraction data, deposition of structures in the Protein Data Bank, and improvement of the quality of deposited crystallographic data.

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-1等其他几个来源的逆转录病毒蛋白酶。已经分析了后一种酶的许多抑制剂复合物，目的是协助开发针对HTLV引起的白血病的药物。蟑螂过敏原BLA G 2被证明是一种非活性天冬氨酸蛋白酶，我们解决了具有不同特异性抗体的两个配合物的结构。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。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。我们已经将Griffithsin重新设计为单体形式，并用复杂的寡糖解决了其结构，从而阐明了其抗病毒特性的基础。我们还解决了另一种凝集素Scytovirin的原子分辨率结构。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。我们已经解决了与其受体复合的Lambda干扰素的结构。晶体学方法论的开发我们一直在研究与衍射数据相相关的问题，蛋白质数据库中结构的沉积以及改善沉积晶体学数据的质量。