Dynamics of transmembrane dimers in TNF-Receptors by EPR and molecular simulation

通过 EPR 和分子模拟研究 TNF 受体跨膜二聚体的动态

• 批准号: 8693092
• 负责人: Jonathan N Sachs
• 金额: $ 34.48万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8693092
• 关键词: Affect; Affinity; Amino Acid Motifs; Amino Acids; Apoptosis; Apoptotic; Architecture; Basic Science; Biological Models; Biophysics; Cancer Intervention; Cancerous; Cell Death; Cell Line; Cells; Cessation of life; Clinical; Collaborations; Complement; Computer Simulation; Coupled; Data; Death Receptor 5; Dimerization; Disease; Drug Design; Electron Spin Resonance Spectroscopy; Event; Extracellular Domain; Extracellular Structure; Family member; Fluorescence; Fluorescence Resonance Energy Transfer; Future; Gene Mutation; Goals; Health; Interdisciplinary Study; Laboratories; Length; Life; Ligand Binding; Ligand Binding Domain; Ligands; Lipids; Liver; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Membrane; Membrane Biology; Membrane Proteins; Modeling; Molecular; Molecular Biology; Molecular Conformation; Mutagenesis; Normal Cell; Paper; Pathway interactions; Play; Process; Proteins; Publishing; Receptor Activation; Receptor Signaling; Recording of previous events; Regulation; Research; Research Personnel; Resolution; Role; Signal Transduction; Structure; Structure-Activity Relationship; Tertiary Protein Structure; Testing; Therapeutic; Therapeutic Intervention; Translating; Transmembrane Domain; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Validation; Vesicle; Work; base; cancer cell; computer studies; dimer; disulfide bond; human disease; innovation; insight; interfacial; member; molecular dynamics; pre-clinical; receptor; receptor function; reconstitution; research study; simulation; therapeutic target

DESCRIPTION (provided by applicant): Death Receptor 5 (DR5) is a member of the TNF-superfamily of transmembrane receptors that plays a critical role in signaling the apoptotic pathway. Upregulated in cancer cells, DR5 is among the most actively pursued anti-cancer targets, both clinically and in basic research studies. However, there is great need to develop new strategies for targeting DR5 in order to maximize apoptosis in cancer cells. To do so, we need significantly more structural and biophysical data in order to understand how the receptor works. Traditionally, research has focused on the crystal structure of the extracellular, ligand-binding domain. As such, there remains a debilitating scarcity of data regarding the key structural events that occur within the transmembrane domain of the protein during transduction of the signal. Very recently, multiple high impact studies have shown that understanding the ligand-induced changes in the structure and dynamics of the transmembrane domain ?- helical dimer is the next crucial step in understanding the function of the receptor. The objective of thi application is to understand key changes in the transmembrane structure of DR5 associated with the active and inactive states of the receptor and to determine the critical amino-acid motifs that dictate changes in conformation. The rationale of this proposal is that once we understand important conformational states of the DR5 TM domain, and the most relevant motifs that stabilize states of the protein, we will be able to evaluate its potential as a therapeutic target or pharmacological regulation. Our approach combines molecular biophysics experiments on model systems (synthetic TM domains) complemented by computational simulations and molecular biology experiments on full-length receptors in living cells. We will 1) define the inter helical architecture of the DR5 transmembrane domain dimer; 2) identify key sequence alterations that either disrupt TM dimerization or stabilize alternate dimer conformations; and 3) establish the capacity to modulate TM dimer architecture and affect DR5 activation in cancer cells. The proposed research will advance understanding of TNF-Receptors in general, taking the logical but critical next steps in building a complete description of their structure-function relationship. We will create new understanding of the physical principles that dictate conformational dynamics of TM dimers, principles that are essential in a broad range of membrane protein superfamilies. In the process, we will advance the state-of-the- art in computational modeling of membrane proteins, providing a methodological roadmap for validation of models by comparison to experimental EPR spectroscopy. Working with a pancreatic cancer researcher will enable us to evaluate the TM domain of DR5 as a target for future therapeutic intervention.

描述（由申请人提供）：死亡受体5（DR 5）是跨膜受体TNF超家族的成员，在细胞凋亡途径的信号传导中起关键作用。在癌细胞中上调，DR 5是临床和基础研究中最积极追求的抗癌靶点之一。然而，非常需要开发靶向DR 5的新策略，以使癌细胞中的凋亡最大化。要做到这一点，我们需要更多的结构和生物物理数据，以了解受体如何工作。传统上，研究集中在胞外配体结合结构域的晶体结构上。因此，在信号转导过程中，关于蛋白质跨膜结构域内发生的关键结构事件的数据仍然缺乏。最近，多个高影响力的研究表明，了解配体诱导的跨膜结构域的结构和动力学变化？螺旋二聚体是了解受体功能的下一个关键步骤。本申请的目的是了解与受体的活性和非活性状态相关的DR 5跨膜结构的关键变化，并确定关键氨基酸基序 决定了构象的变化。这个建议的基本原理是，一旦我们了解DR 5 TM结构域的重要构象状态，以及稳定蛋白质状态的最相关基序，我们将能够评估其作为治疗靶点或药理学调节的潜力。我们的方法结合了分子生物物理学实验的模型系统（合成TM域）的计算模拟和分子生物学实验的全长受体在活细胞中的补充。我们将1）定义内部 DR 5跨膜结构域二聚体的螺旋结构; 2）鉴定破坏TM二聚化或稳定替代二聚体构象的关键序列改变;和3）建立调节TM二聚体结构和影响癌细胞中DR 5活化的能力。拟议的研究将促进对TNF受体的总体理解，采取逻辑但关键的下一步，建立其结构-功能关系的完整描述。我们将创造新的理解的物理原理，决定TM二聚体的构象动力学，原则是必不可少的，在广泛的膜蛋白超家族。在这个过程中，我们将推进膜蛋白计算建模的最新技术，通过与实验EPR光谱的比较，为模型的验证提供方法路线图。与胰腺癌研究人员合作将使我们能够评估DR 5的TM结构域作为未来治疗干预的靶点。