Effects of lipidomic diversity on GPCR

脂质组多样性对 GPCR 的影响

基本信息

批准号：
10246266
负责人：
Edward Ray Lyman
金额：
$ 30.39万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10246266
关键词：
ADORA2A gene Affect Binding Proteins Biochemical Biological Assay Biological Models Biophysics Blood flow Cardiac Myocytes Cell Culture Techniques Cell Membrane Proteins Cell membrane Cells Cellular biology Characteristics Cholesterol Complex Complex Mixtures Computer Models Controlled Environment Coupling Cultured Cells Data Dependence Diet Dietary Fats Environment Epithelial Cells Fatty Acids Follow-Up Studies G-Protein-Coupled Receptors GTP-Binding Proteins Genetic Goals Heart Individual Integral Membrane Protein Kinetics Knowledge Leukocytes Ligand Binding Ligands Lipid Bilayers Lipids Lovastatin Measurement Measures Membrane Membrane Proteins Molecular Molecular Conformation Mutagenesis Neurons Parkinson Disease Pharmaceutical Preparations Pharmacologic Substance Phenotype Physiological Plasma Cells Property Proteins Purinergic P1 Receptors Receptor Signaling Records Research Signal Transduction Site Specificity Sterols Stress Testing Therapeutic Thermodynamics Tissues Variant Viscosity base cell type cellular imaging feeding inhibitor/antagonist lipidome lipidomics membrane model methyl-beta-cyclodextrin personalized medicine physical property protein function proteoliposomes prototype receptor receptor function reconstitution response simulation targeted treatment therapeutic target unilamellar vesicle

项目摘要

Integral membrane protein (IMP) function depends on the membrane environment, both through the activity of specific lipids as allosteric modulators and via bulk membrane properties like viscosity, rigidity, and order. Recent lipidomic analyses have established the compositional details of membrane complexity, and provided clear evidence that disparate cell types — and even the same cell type in different individuals — present unique, grossly different membrane environments. Our preliminary observations suggest that these distinct membrane environments influence the conformation and activity of G protein-coupled receptors (GPCRs), and likely modulate their response to targeted therapeutics. Moreover, our recent observations reveal that membrane phenotypes are remarkably susceptible to exogenous perturbations, suggesting that extrinsic factors like diet or lipid synthesis inhibitors could synergize with GPCR-targeted therapeutics. It is therefore critical to define the spectrum of regulatory mechanisms imparted on proteins by lipid bilayers, with this knowledge especially impactful for high value targets, such as GPCRs. Our long-term goal is to exploit tissue-specific differences in membrane environments to predict and tune the efficacy and specificity of therapeutics targeting GPCRs. The objective of this proposal is to leverage advances in lipidomics and GPCR reconstitution to bridge the membrane complexity of live cells with the controlled environments provided by model systems. By combining genetics, cell biology, lipidomics, GPCR reconstitution into proteoliposomes, and molecular simulations we aim to determine the mechanisms by which membrane composition determines ligand binding and signaling characteristics for the adenosine A2A receptor (A2AR). Our central hypothesis is that ligand off-rates and G protein coupling are affected by local membrane order, which is a function of both cholesterol concentration and overall lipid composition. Across model systems, our preliminary observations suggest that cholesterol concentration is a key modulator of GPCR activity, with effects on both ligand binding thermodynamics and signaling kinetics. We will decipher whether the effects of cholesterol are due to specific interactions with the receptor or effects on membrane physical properties by independently varying and measuring these properties in simulations, model membranes, and live cells. Lipidomic analysis of A2AR-relevant target cell types — neurons, leukocytes, cardiomyocytes, and epithelial cells — will inform molecular simulations of complex mixtures, and reconstitution of A2AR into membranes obtained from these same cells will determine the extent of signaling variation across cell types. Finally, we will test whether the effects of lipidomic perturbations — including cholesterol depletion / loading, fatty acid feeding, and drugs that modulate fatty acid and cholesterol synthesis — recapitulate A2AR functional dependence observed in model systems. This final goal will test a prototype therapy based on lipidomic perturbation, to locally and specifically enhance A2AR therapeutics..

整体膜蛋白（IMP）功能取决于膜环境特定的脂质作为变构调节剂，并通过散装膜特性（例如粘度，刚度和顺序）。最近的脂质组分析已经建立了膜复杂性的综合细节，并提供了清晰的证据表明不同个体不同细胞类型（甚至在不同个体中的相同细胞类型）都具有独特的证据严重不同的膜环境。我们的初步观察表明这些独特的膜环境影响G蛋白偶联受体（GPCR）的会议和活动，并且可能调节他们对靶向治疗的反应。此外，我们最近的观察结果表明膜表型非常容易受到外源性扰动的影响，表明饮食或脂质合成抑制剂可以与靶向GPCR的疗法协同作用。因此，定义脂质双层赋予蛋白质的调节机制的光谱，尤其是这些知识对高价值目标（例如GPCR）的影响。我们的长期目标是探索膜环境中组织特异性差异以预测和调整针对GPCR的治疗疗法的效率和特异性。该提议的目的是利用脂质组学和GPCR重构的进步，桥接活细胞的膜复杂性模型系统提供的控制环境。通过结合遗传学，细胞生物学，脂肪态学，GPCR 重组为蛋白质脂质体和分子模拟，我们旨在确定该机制膜组成确定腺苷A2A接收器的配体结合和信号传导特性（A2AR）。我们的中心假设是配体离子率和G蛋白偶联受局部影响膜序，这是胆固醇浓度和整体脂质组成的函数。在整个模型系统中，我们的初步观察结果表明胆固醇浓度是关键 GPCR活性的调节剂，对配体结合热力学和信号动力学的影响。我们将破译胆固醇的作用是由于与接收器的特定相互作用或对膜物理特性通过独立变化和测量模拟中的这些特性，模型膜和活细胞。 A2AR相关靶细胞类型的脂质组分析 - 神经元，白细胞，心肌细胞和上皮细胞 - 将告知复杂混合物的分子模拟，并重新建立从这些相同细胞获得的A2AR进入膜的膜将决定跨这些信号变化的程度细胞类型。最后，我们将测试脂肪组扰动的影响 - 包括胆固醇耗竭 / 负载，脂肪酸进食以及调节脂肪酸和胆固醇合成的药物 - 概括A2AR 在模型系统中观察到的功能依赖性。这个最终目标将测试基于脂质组的原型疗法扰动，以局部和专门增强A2AR治疗。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Varying the position of phospholipid acyl chain unsaturation modulates hopanoid and sterol ordering.

改变磷脂酰基链不饱和度的位置可调节霍帕尼和甾醇的排序。

DOI：
10.1101/2023.09.06.556521
发表时间：
2024
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Nguyen,Ha-Ngoc-Anh;Sharp,Liam;Lyman,Edward;Saenz,JamesP
通讯作者：
Saenz,JamesP

Ultrafast Formation of the Charge Transfer State of Prodan Reveals Unique Aspects of the Chromophore Environment.

DOI：
10.1021/acs.jpcb.0c00121
发表时间：
2020-04-02
期刊：
The journal of physical chemistry. B
影响因子：
0
作者：
Baral S;Phillips M;Yan H;Avenso J;Gundlach L;Baumeier B;Lyman E
通讯作者：
Lyman E

Lipid-Protein Interactions in Plasma Membrane Organization and Function.

质膜组织和功能中的脂质-蛋白质相互作用。

DOI：
10.1146/annurev-biophys-090721-072718
发表时间：
2022
期刊：
Annual review of biophysics
影响因子：
12.4
作者：
Sych,Taras;Levental,KandiceR;Sezgin,Erdinc
通讯作者：
Sezgin,Erdinc

Curvature Energetics Determined by Alchemical Simulation on Four Topologically Distinct Lipid Phases.

DOI：
10.1021/acs.jpcb.0c09458
发表时间：
2021-02-25
期刊：
JOURNAL OF PHYSICAL CHEMISTRY B
影响因子：
3.3
作者：
Beaven, Andrew H.;Arnarez, Clement;Lyman, Edward;Bennett, W. F. Drew;Sodt, Alexander J.
通讯作者：
Sodt, Alexander J.

Surface Shear Viscosity and Interleaflet Friction from Nonequilibrium Simulations of Lipid Bilayers.

脂质双层非平衡模拟的表面剪切粘度和叶间摩擦。