The a-catulin/dystrophin-associated protein complex regulates GPCR function.

α-catulin/肌营养不良蛋白相关蛋白复合物调节 GPCR 功能。

• 批准号: 9380078
• 负责人: Chris S Hague
• 金额: $ 34.69万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-07 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9380078
• 关键词: Adrenergic Agents; Adrenergic Antagonists; Adrenergic Receptor; Adrenergic alpha-Antagonists; Affect; Afghanistan; Antihypertensive Agents; Architecture; Benign Prostatic Hypertrophy; Binding; Biochemical; Biochemical Process; Biological Assay; Biology; Blood Vessels; Cardiovascular Diseases; Catecholamines; Cell Culture Techniques; Cell membrane; Cell model; Cell physiology; Cells; Co-Immunoprecipitations; Colon Carcinoma; Complex; Contracts; Cultured Cells; DLG1 gene; DLG4 gene; Development; Disease; Drug usage; Dystrophin-Associated Protein Complex; Elderly; Endoplasmic Reticulum; Ensure; Environment; Funding; G-Protein-Coupled Receptors; Genes; Goals; Human; Human Cell Line; Hypertension; Image; In Situ; In Vitro; Iraq; Knowledge; LPAR4 gene; Ligands; Link; Lipids; Malignant Epithelial Cell; Mass Spectrum Analysis; Modeling; Molecular; Morbidity - disease rate; Motor Activity; Myocardial Infarction; Neuraxis; Nightmare; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Post-Traumatic Stress Disorders; Prazosin; Prostate; Proteins; Proteomics; Receptor Signaling; Role; SW480; Safety; Schizophrenia; Series; Shapes; Signal Transduction; Stenosis; Stimulus; Sympathetic Nervous System; System; Technology; Testing; Toxic effect; Urinary Incontinence; Urine; Veterans; Western Blotting; blood pressure regulation; brain volume; combat; fighting; insight; male; novel; polarized cell; prevent; protein complex; radioligand; receptor; receptor function; small molecule; success; syntrophin; theories; therapeutic target

Abstract: α1D-adrenergic receptors (ARs) are essential G protein-coupled receptors (GPCRs) of the sympathetic nervous system, and are a promising therapeutic target for an array of diseases. In the central nervous system, the α1D-AR tightly regulates stimulus-induced locomotor activity, and is 1 of 13 hypermethylated genes associated with decreased brain volume in schizophrenic patients. The α1D-AR is critical for blood pressure regulation and stenosis of damaged blood vessels. The receptor also can negatively impact urine flow by contracting the prostate in patients suffering from benign prostatic hypertrophy (BPH). Thus, α1-AR antagonists (“α-blockers”) are often used to treat hypertension, urinary incontinence, and most recently with promising success stories, to prevent reoccurring nightmares in combat veterans afflicted with Post-Traumatic Stress Disorder. Unfortunately, major toxicities can often occur in patients taking α-blockers. During the ALLHAT trial, α-blocker therapy was discontinued due to increased patient morbidity. Accordingly, a clearer picture of how the α1D-AR engages with its cellular environment will provide critical insights towards the further development of small molecule α1D-AR modulators beneficial for the treatment of PTSD, BPH, and cardiovascular disease. Surprisingly, our basic knowledge of α1D-AR biochemical processes is lacking within human contexts, primarily because no human cell lines have been identified that express endogenous α1D-ARs. Without adequate cell culture models and human model cell systems to examine their discrete biochemical interactions, it will continue to be challenging to develop new small molecules targeting α1D-ARs and to understand their essential molecular and cellular functions. We have made significant progress towards solving some of these mysteries. First, we discovered that α1D-ARs interact with multiple PSD95/DLG1/Zo-1 (PDZ) domain-containing proteins. Second, we found these interactions are essential for α1D-ARs to be expressed as functional receptors at the plasma membrane. Remarkably, we found that α1D-ARs interact with two PDZ- proteins, syntrophin and scribble, in all human cell lines we examined. This novel discovery provides an opportunity to develop small molecule allosteric ligands targeting α1D-AR:PDZ-protein interaction-interfaces. However, this first requires a thorough characterization of α1D-AR:PDZ-protein architecture and function. In this competing renewal, we propose to extend our findings in new directions using the following aims. Aim 1: Determine whether scribble organizes α1D-ARs into signaling clusters. Aim 2: Identify α1D-AR:PDZ-protein complex(es) in human cells. This proposal has the overarching goal of targeting discrete α1D-AR:PDZ-protein interfaces with novel small molecules to treat diseases associated with aberrant α1D-AR signaling.

翻译后摘要：α 1D-肾上腺素能受体（AR）是必不可少的G蛋白偶联受体（GPCRs）的 交感神经系统，并且是一系列疾病的有希望的治疗靶点。中部 在神经系统中，α1D-AR紧密调节刺激诱导的运动活动，是13种中的1种。 与精神分裂症患者脑容量减少相关的高甲基化基因α1D-AR是 对于血压调节和受损血管的狭窄至关重要。受体也可以负性地 通过收缩患有良性前列腺肥大（BPH）的患者的前列腺来影响尿流。 因此，α1-AR拮抗剂（“α-阻断剂”）通常用于治疗高血压、尿失禁和大多数糖尿病。 最近有一些有希望的成功故事，以防止受战争折磨的退伍军人再次出现噩梦。 创伤后应激障碍。不幸的是，服用α受体阻滞剂的患者通常会发生严重的毒性反应。 在ALLHAT试验期间，由于患者发病率增加而停止α受体阻滞剂治疗。因此 更清楚地了解α1D-AR如何与其细胞环境结合，将为研究α1D-AR的作用提供重要的见解。 进一步开发有益于治疗PTSD、BPH的小分子α1D-AR调节剂， 心血管疾病令人惊讶的是，我们对α1D-AR生化过程的基本知识是缺乏的， 人类背景，主要是因为尚未鉴定出表达内源性α 1D-AR的人类细胞系。 没有足够的细胞培养模型和人类模型细胞系统来检查它们的离散生化 因此，开发新的靶向α 1D-AR的小分子和 了解它们的基本分子和细胞功能。我们已经在解决 其中的一些谜团。首先，我们发现α 1D-AR与多个PSD 95/DLG 1/Zo-1（PDZ）相互作用， 含结构域的蛋白质。其次，我们发现这些相互作用对于α 1D-AR表达为 质膜上的功能性受体。值得注意的是，我们发现α 1D-AR与两个PDZ相互作用， 蛋白质，syntrophin和scribble，在我们检测的所有人类细胞系中。这一新发现提供了一个 有机会开发靶向α1D-AR的小分子变构配体：PDZ-蛋白质相互作用界面。 然而，这首先需要彻底表征α1D-AR：PDZ蛋白的结构和功能。在这 为了竞争更新，我们建议使用以下目标将我们的研究结果扩展到新的方向。 目的1：确定scribble是否将α 1D-AR组织成信号簇。 目的2：鉴定人细胞中α1D-AR：PDZ蛋白复合物。 该提案的总体目标是靶向离散的α1D-AR：PDZ-蛋白质界面， 分子治疗与异常α1D-AR信号传导相关的疾病。