Streamlined Structures of Human Integral Membrane Proteins at Atomic Resolution

原子分辨率下人类整体膜蛋白的简化结构

• 批准号: 8146511
• 负责人: STEPHEN G ALLER
• 金额: $ 219.75万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8146511
• 关键词: Acquired Immunodeficiency Syndrome; Affinity; Amino Acids; Antibodies; Antigens; Binding; Carrier Proteins; Communication; Crystallography; Cystic Fibrosis; Diabetes Mellitus; Disease; Drug Delivery Systems; Drug Design; Engineering; Human; Human Genome; Inflammation; Integral Membrane Protein; Libraries; Light; Malignant Neoplasms; Methods; Molecular; Molecular Chaperones; Molecular Conformation; Multi-Drug Resistance; Pharmaceutical Preparations; Polycystic Kidney Diseases; Process; Production; Proteins; Resolution; Role; Screening procedure; Structural Biochemistry; Structure; System; Time; Yeasts; absorption; abstracting; base; cost; drug discovery; flexibility; innovation; protein structure; public health relevance; scaffold; three dimensional structure

DESCRIPTION (Provided by the applicant) Abstract: Streamlined Structures of Human Integral Membrane Proteins at Atomic Resolution About 35% of the human genome encodes integral membrane proteins (IMPs) and one-third of approved drugs target this class. While human IMPs are amenable to straightforward biochemistry, structural studies are nearly intractable as evidenced by the paucity of human IMP structures available. To date, only five human IMP structures have been solved to truly atomic resolution (< 3.0 ¿). None of the human IMP structures represent transporters, likely due to the extreme flexibility from alternating access mechanisms impeding crystallography. Sharp resolution of IMPs in multiple conformations is a prerequisite for understanding the full mechanism of transport proteins, the role of amino acids in substrate recognition, drug binding, inter-domain communication and for accurate structure-based drug design. Structure-determination of human IMPs lags decades behind the determination of soluble protein structures. We plan to accelerate the process and simultaneously provide human IMP structures in multiple conformations at atomic resolution. Our innovative strategy utilizes the screening of a synthetic antibody library to rapidly identify high- affinity Fabs (SynFabs) that will serve as scaffolds for crystallography. The versatile SynFab library can recognize virtually limitless numbers of antigens, will trap human IMPs in multiple conformations and will universalize the structure determination process using molecular replacement methods. We have engineered molecular chaperones to enable the production of the most difficult human IMPs in folded mature form using a low-cost yeast expression system. Our innovative and comprehensive strategy will accelerate structure determination of human IMPs by years, shedding light on the mechanisms of serious diseases including Cystic Fibrosis (CF), diabetes, cancer, polycystic kidney disease, inflammation, AIDS and multi-drug resistance (MDR). Public Health Relevance: Many diseases are directly caused by a major class of proteins called integral membrane proteins (IMPs). Obtaining three-dimensional structures of human IMPs has been far too costly and time-consuming for effective drug design or the computational prediction of drug absorption, permeation of drug barriers, and multi-drug resistance. We employ a new strategy to determine structures of human IMPs at highthroughput to integrate these computational approaches and accelerate drug discovery.

描述（由申请人提供） 摘要：原子分辨率下人类整合膜蛋白的简化结构大约 35% 的人类基因组编码整合膜蛋白 (IMP)，三分之一的已批准药物针对此类蛋白。虽然人类 IMP 适合简单的生物化学研究，但结构研究几乎是棘手的，可用的人类 IMP 结构的缺乏就证明了这一点。迄今为止，只有五个人类 IMP 结构被解析到真正的原子分辨率（< 3.0 ¿）。人类 IMP 结构中没有一个代表转运蛋白，这可能是由于交替访问机制的极大灵活性阻碍了晶体学。多种构象中 IMP 的清晰分辨率是了解转运蛋白的完整机制、氨基酸在底物识别、药物结合、域间通信中的作用以及基于结构的精确药物设计的先决条件。人类 IMP 的结构测定比可溶性蛋白质结构的测定落后数十年。我们计划加速这一过程，同时以原子分辨率提供多种构象的人类 IMP 结构。我们的创新策略利用合成抗体库的筛选来快速识别高亲和力 Fab (SynFab)，将其用作晶体学支架。多功能的 SynFab 文库可以识别几乎无限数量的抗原，将人类 IMP 捕获为多种构象，并将使用分子替换方法普遍化结构测定过程。我们设计了分子伴侣，能够使用低成本酵母表达系统以折叠成熟形式生产最困难的人类 IMP。我们创新而全面的策略将加速人类 IMP 的结构测定，揭示囊性纤维化 (CF)、糖尿病、癌症、多囊肾病、炎症、艾滋病和多重耐药性 (MDR) 等严重疾病的机制。 公共卫生相关性：许多疾病是由称为整合膜蛋白 (IMP) 的主要蛋白质类别直接引起的。对于有效的药物设计或药物吸收、药物屏障渗透和多药耐药性的计算预测来说，获得人类 IMP 的三维结构成本太高且耗时。我们采用一种新策略来确定高通量的人类 IMP 结构，以整合这些计算方法并加速药物发现。