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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

基本信息

批准号：
9250185
负责人：
Jonathan N Sachs
金额：
$ 33.18万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9250185
关键词：
Affect Affinity Amino Acid Motifs Amino Acids Apoptosis Apoptotic Architecture Basic Science Biological Models Biology Biophysics Cancer Intervention Cancerous Cell Line Cell model Cells Cessation of life Clinical Collaborations Complement Computer Simulation Coupled Crystallization DNA Sequence Alteration Data Death Domain Death Receptor 5 Dimerization Disease Drug Design Electron Spin Resonance Spectroscopy Event Extracellular Domain Extracellular Structure Family member Fluorescence Fluorescence Resonance Energy Transfer Future Goals Interdisciplinary Study Laboratories Length Ligand Binding Ligand Binding Domain Ligands Lipids Liver Malignant Neoplasms Malignant neoplasm of pancreas Measures Membrane Membrane Proteins Methodology Modeling Molecular Molecular Biology Molecular Computations Molecular Conformation Mutagenesis Normal Cell Paper Pathway interactions Pharmacology Play Process Proteins Publishing Receptor Activation Receptor Signaling Recording of previous events Regulation Research Research Personnel Resolution Role Signal Transduction Structure Structure-Activity Relationship TNF gene Tertiary Protein Structure Testing Therapeutic Therapeutic Intervention Translating Transmembrane Domain Tumor Necrosis Factor Receptor Validation Vesicle Work alpha helix base cancer cell computer studies dimer disulfide bond experimental study human disease innovation insight interfacial member molecular dynamics pre-clinical public health relevance receptor receptor function reconstitution research study simulation therapeutic target

项目摘要

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 α-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(申请人提供)：死亡受体5(DR5)是肿瘤坏死因子超家族跨膜受体中的一员，在细胞凋亡途径中起着关键作用。DR5在癌细胞中上调，在临床和基础研究中都是最积极追求的抗癌靶点之一。然而，为了最大限度地促进癌细胞的凋亡，迫切需要开发新的靶向DR5的策略。要做到这一点，我们需要更多的结构和生物物理数据来了解受体是如何工作的。传统上，研究主要集中在细胞外配体结合域的晶体结构上。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很少。最近，多项高影响力的研究表明，了解配体诱导的跨膜区α-螺旋二聚体的结构和动力学变化是理解受体功能的下一个关键步骤。本应用的目的是了解DR5跨膜结构中与受体活性和非活性状态相关的关键变化，并确定决定构象变化的关键氨基酸基序。这一建议的基本原理是，一旦我们了解了DR5TM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物调节的潜力。我们的方法结合了模型系统(合成TM结构域)上的分子生物物理实验，以及活细胞中全长受体的计算模拟和分子生物学实验。我们将1)确定DR5跨膜结构域二聚体的螺旋间结构；2)确定破坏TM二聚体或稳定交替二聚体构象的关键序列改变；以及3)建立调节TM二聚体结构并影响癌细胞中DR5激活的能力。这项拟议的研究将在总体上促进对肿瘤坏死因子受体的理解，采取合乎逻辑但关键的下一步，以建立对其结构-功能关系的完整描述。我们将对决定TM二聚体构象动力学的物理原理产生新的理解，这些原理在广泛的膜蛋白超家族中是必不可少的。在这个过程中，我们将推进膜蛋白计算建模的最新进展，通过与实验EPR波谱的比较，为模型的验证提供方法学路线图。与胰腺癌研究人员合作将使我们能够评估DR5的TM结构域，作为未来治疗干预的目标。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Death Receptor 5 Activation Is Energetically Coupled to Opening of the Transmembrane Domain Dimer.

死亡受体 5 的激活与跨膜域二聚体的打开呈能量耦合。

DOI：
10.1016/j.bpj.2017.05.038
发表时间：
2017
期刊：
Biophysical journal
影响因子：
3.4
作者：
Vunnam,Nagamani;Campbell-Bezat,CecilyKristine;Lewis,AndrewK;Sachs,JonathanN
通讯作者：
Sachs,JonathanN

Piecing it together: Unraveling the elusive structure-function relationship in single-pass membrane receptors.

DOI：
10.1016/j.bbamem.2017.01.016
发表时间：
2017-09
期刊：
Biochimica et biophysica acta. Biomembranes
影响因子：
0
作者：
Valley CC;Lewis AK;Sachs JN
通讯作者：
Sachs JN

HuR Contributes to TRAIL Resistance by Restricting Death Receptor 4 Expression in Pancreatic Cancer Cells.

DOI：
10.1158/1541-7786.mcr-15-0448
发表时间：
2016-07
期刊：
Molecular cancer research : MCR
影响因子：
0
作者：
Romeo C;Weber MC;Zarei M;DeCicco D;Chand SN;Lobo AD;Winter JM;Sawicki JA;Sachs JN;Meisner-Kober N;Yeo CJ;Vadigepalli R;Tykocinski ML;Brody JR
通讯作者：
Brody JR

Death Receptor 5 Networks Require Membrane Cholesterol for Proper Structure and Function.

死亡受体 5 网络需要膜胆固醇才能实现适当的结构和功能。

DOI：
10.1016/j.jmb.2016.10.001
发表时间：
2016
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Lewis,AndrewK;Valley,ChristopherC;Peery,StephenL;Brummel,Benjamin;Braun,AnthonyR;Karim,ChristineB;Sachs,JonathanN
通讯作者：
Sachs,JonathanN

Soluble Extracellular Domain of Death Receptor 5 Inhibits TRAIL-Induced Apoptosis by Disrupting Receptor-Receptor Interactions.

死亡受体 5 的可溶性胞外结构域通过破坏受体-受体相互作用来抑制 TRAIL 诱导的细胞凋亡。