MLP Assay for Arrestin-AP2 Inhibitors

Arrestin-AP2 抑制剂的 MLP 测定

• 批准号: 8208096
• 负责人: Eric R Prossnitz
• 金额: $ 3.78万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8208096
• 关键词: Adaptor Signaling Protein; Apoptosis; Apoptotic; Arrestins; Binding; Biological; Biological Assay; Capsid Proteins; Cells; Clathrin; Collection; Disease; Down-Regulation; Ear; Endosomes; Family; Flow Cytometry; Funding; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Glutathione; Goals; Human; Human Biology; Left; Ligand Binding; Mediating; Mitogen-Activated Protein Kinases; Molecular; Molecular Probes; New Mexico; Pathway interactions; Peptides; Phosphotransferases; Physiology; Play; Proteins; Publications; Receptor Activation; Recycling; Regulation; Research; Research Personnel; Role; SRC gene; Signal Pathway; Signal Transduction; Signaling Protein; System; Therapeutic; Transcription Factor AP-2 Alpha; Universities; Vesicle; arrestin 2; base; cancer cell; desensitization; experience; fMet-Leu-Phe receptor; formyl peptide; human disease; inhibitor/antagonist; novel; prevent; public health relevance; receptor; receptor binding; receptor function; receptor internalization; small molecule; therapeutic development; tool; trafficking

DESCRIPTION (provided by applicant): G protein-coupled receptors (GPCR) play a critical role in almost every aspect of human physiology and disease. The protein arrestin plays a vital role in many of these pathways. Arrestin binds to the phosphorylated form of GPCRs and prevents the receptor from binding to and activating G proteins, resulting in desensitization. In addition, for many GPCRs, arrestin functions as an adapter molecule for internalization as well as "secondary" signaling. Arrestin mediates internalization via a domain that binds directly to clathrin and the adapter AP-2. Arrestins also bind to kinases and other signaling proteins. Our recent results have identified a specific molecular interaction between arrestin and AP-2 that is critical for the recycling certain GPCRs. We have also shown that disrupting this interaction induces a rapid apoptotic pathway for many GPCRs. To date there are no small molecules known that regulate interactions between arrestin and its interacting proteins. We have developed a high throughput flow cytometric assay involving AP-2 and a fluorescent arrestin peptide. The primary goal of the proposed research is to identify compounds that regulate the interaction between arrestin and AP-2, resulting in modulation of GPCR trafficking, signaling and apoptosis. A small molecule that inhibits this interaction will be a valuable tool to assess the role of AP-2 in the arrestin-dependent regulation of potentially hundreds of GPCRs and could represent the basis for therapeutic development through its ability to induce apoptosis in disease cells. PUBLIC HEALTH RELEVANCE: Receptors play a vital role in almost every aspect of human biology and disease. Receptor activity and function are mediated by a large collection of interacting proteins. The largest class of receptors is the G protein-coupled receptor (GPCR) family. The protein arrestin plays a critical role in both inhibiting and activating signaling by GPCRs. We have identified a specific interaction between arrestin and the cellular protein AP-2. To date there are no small molecules known that regulate interactions between arrestin and its interacting proteins. The goal of the proposed research is to identify compounds that inhibit or stimulate the interaction between arrestin and AP-2 to provide a valuable tool to assess the role of this interaction in the regulation of hundreds of GPCRs. Such a molecule would be of great benefit for researchers in the field and could find therapeutic utility in its ability to induce apoptosis in hyper stimulated disease cells such as cancer cells.

描述（由申请人提供）：G蛋白偶联受体（GPCR）在人类生理和疾病的几乎每个方面都起着至关重要的作用。蛋白阻滞素在许多此类途径中起着至关重要的作用。阻止蛋白与GPCR的磷酸化形式结合，并防止受体与和激活G蛋白结合并激活G蛋白，从而导致脱敏。此外，对于许多GPCR，逮捕蛋白作为内在化和“次级”信号传导的适配器分子起作用。逮捕蛋白通过直接与网格蛋白和适配器AP-2结合的结构域介导内在化。逮捕蛋白还与激酶和其他信号蛋白结合。我们最近的结果已经确定了抑制蛋白和AP-2之间的特定分子相互作用，这对于回收某些GPCR至关重要。我们还表明，破坏这种相互作用会引起许多GPCR的快速凋亡途径。迄今为止，还没有知道抑制蛋白与其相互作用蛋白之间相互作用的小分子。我们已经开发了涉及AP-2和荧光阻止肽肽的高通量流式细胞仪测定。拟议研究的主要目标是鉴定调节抑制蛋白和AP-2之间相互作用的化合物，从而导致GPCR贩运，信号传导和凋亡的调节。一种抑制这种相互作用的小分子将是评估AP-2在依赖抑制蛋白依赖数百GPCR中的作用的有价值的工具，并且可以通过其在疾病细胞中诱导凋亡的能力来代表治疗性发育的基础。 公共卫生相关性：受体在人类生物学和疾病的几乎每个方面都起着至关重要的作用。受体活性和功能由大量相互作用的蛋白质介导。最大的受体类是G蛋白偶联受体（GPCR）家族。蛋白阻滞蛋白在GPCR抑制和激活信号传导中起着至关重要的作用。我们已经确定了抑制蛋白和细胞蛋白AP-2之间的特定相互作用。迄今为止，还没有知道抑制蛋白与其相互作用蛋白之间相互作用的小分子。拟议的研究的目的是确定抑制或刺激抑制蛋白和AP-2之间相互作用的化合物，以提供有价值的工具来评估这种相互作用在数百种GPCR中的作用。这样的分子对于该领域的研究人员会带来很大的好处，并且可以在诱导超刺激性疾病细胞（例如癌细胞）中凋亡的能力找到治疗效用。