Understanding the structural dynamics of TNF receptors

了解 TNF 受体的结构动力学

• 批准号: 10178044
• 负责人: Jonathan N Sachs
• 金额: $ 37.35万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10178044
• 关键词: Address; Apoptosis; Aromatic Amino Acids; Autoimmune Diseases; Binding; Biophysics; Clinical; Disease; Extracellular Domain; Fluorescence Resonance Energy Transfer; Goals; Grant; In Vitro; Industry; Inflammatory; Integral Membrane Protein; Journals; Ligand Binding; Malignant Neoplasms; Membrane; Methionine; Methods; Modeling; Molecular Computations; Molecular Conformation; Pharmaceutical Preparations; Play; Process; Productivity; Proteins; Publishing; Receptor Activation; Receptor Signaling; Research; Rheumatoid Arthritis; Role; Set protein; Signal Transduction; Specificity; Structure; TNFRSF10B gene; TNFRSF1A gene; Techniques; Therapeutic Intervention; Thermodynamics; Time; Transmembrane Domain; Tumor Necrosis Factor Receptor; Vertebral column; Work; anti-cancer; base; biophysical tools; clinically relevant; conformational alteration; drug discovery; experimental study; flexibility; high risk; interdisciplinary collaboration; member; molecular modeling; oxidation; prevent; protein folding; protein misfolding; receptor; side effect; small molecule; therapeutic target; tool

ABSTRACT The tumor necrosis factor receptors (TNFRs) are a superfamily of transmembrane proteins that play critical roles in apoptosis and inflammatory diseases and are considered important therapeutic targets. Even though targeting of TNFRs is a billion-dollar industry, the clinically available drugs cause devastating side effects because they lack receptor specificity. My research focuses on understanding the essential conformational dynamics of TNFRs that transduce signals across the membrane, with the ultimate goal of enabling highly effective and specific targeting. To accelerate scientific discovery, we have focused on two of the most clinically relevant members of the superfamily: TNFR1, involved in various autoimmune diseases, including rheumatoid arthritis; and Death Receptor 5, one of the most actively pursued anti-cancer targets. We apply an investigative strategy that includes computational molecular modeling, thermodynamic calculations, and in vitro experimental tools, enabling us to predict and understand conformational changes in these single-pass transmembrane proteins. Our work has yielded important findings published in high-impact journals. For instance, we elucidated mechanisms of ligand binding in both TNFR1 and DR5. We found that binding is controlled by an interaction between methionine and aromatic amino acids, causing a conformational rearrangement of the ligand-binding pocket. Our studies of this interaction motif led to a fundamental discovery that answered a long-standing question regarding the role of methionine in protein folding, and further, how methionine oxidation causes protein misfolding. We built a new model of TNFR oligomerization that led us to discover that ligand binding causes a large-scale backbone conformational change in the extracellular domain of the receptor. This finding revised previous assumptions regarding TNFRs that activation occurs without any conformational changes in the receptor backbone. With computation and biophysical and cellular experiments, we also showed for the first time a scissors-like opening that occurs in the transmembrane domain helices and explained the fundamental thermodynamics of this process. Significantly, using FRET-based small molecule discovery, we built on our new model of TNFR activation and showed that allosteric alteration of the conformational states of TNFRs can inhibit activation, and have thereby opened new avenues to therapeutic intervention. We propose to extend our discoveries by integrating the dynamic modes across domains of the receptor and answering the fundamental question: what is the structural and dynamic mechanism of TNFR activation? We will address impactful questions, some of which may be high-risk, but with potential to be transformative in the field and to launch new directions in drug discovery. Our productivity is enhanced by longstanding interdisciplinary collaborations that engage additional biophysical tools, including EPR and NMR. The MIRA grant will provide flexibility to methodically, and deeply, address fundamental questions regarding TNFR signaling, which will profoundly enhance efforts to identify and rationally target the most vulnerable structural motifs in these important proteins.!

抽象的 肿瘤坏死因子受体（TNFR）是跨膜蛋白的超家族，起着关键作用 在凋亡和炎症性疾病中，被认为是重要的治疗靶标。即使针对目标 TNFRS是一个十亿美元的行业，临床上可用的药物会引起毁灭性的副作用，因为它们 缺乏受体特异性。我的研究重点是理解TNFR的基本构象动态 该信号在整个膜上传递，其最终目标是使高效和特定 定位。为了加速科学发现，我们专注于两个最相关的成员 超家族：TNFR1，涉及各种自身免疫性疾病，包括类风湿关节炎；和死亡 受体5，最积极的抗癌目标之一。我们采用了一个调查策略，包括 计算分子建模，热力学计算和体外实验工具，使我们能够 预测和理解这些单通跨膜蛋白中的构象变化。我们的工作有 产生了在高影响期刊上发表的重要发现。例如，我们阐明了配体的机制 在TNFR1和DR5中结合。我们发现结合是由蛋氨酸和 芳香氨基酸，导致配体结合口袋的构象重排。我们对此的研究 互动主题导致了一个基本发现，该发现回答了一个关于角色的长期问题 蛋白质折叠中的蛋氨酸，进一步，蛋氨酸氧化如何导致蛋白质错误折叠。我们建造了一个新的 TNFR寡聚的模型，使我们发现配体结合会导致大规模主链 受体细胞外域的构象变化。这一发现修订了以前的假设 关于激活发生的TNFR，没有受体主链没有任何构象变化。和 计算以及生物物理和细胞实验，我们还首次展示了剪刀样开口 这发生在跨膜域螺旋中，并解释了这一点的基本热力学 过程。值得注意的是，使用基于FRET的小分子发现，我们建立在TNFR的新模型上 激活并表明TNFR构象状态的变构改变可以抑制激活，并且 因此，是否为治疗干预开辟了新的途径。我们建议通过 整合跨受体领域的动态模式并回答基本问题：什么 TNFR激活的结构和动态机制是吗？我们将解决有影响力的问题，其中一些 可能是高风险 发现。长期以来的跨学科合作可以提高我们的生产力 生物物理工具，包括EPR和NMR。 Mira Grant将为有条不紊，深刻地提供灵活性 解决有关TNFR信号传导的基本问题，该问题将极大地加强识别和 合理地针对这些重要蛋白质中最脆弱的结构基序。