ER-to-Golgi traffic and signal regulation of GPCRs

GPCR 的 ER 至高尔基体交通和信号调节

• 批准号: 7227755
• 负责人: GUANGYU WU
• 金额: $ 24.2万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7227755
• 关键词: Adrenergic Agents; Angiotensins; Cell Line; Cell Surface Receptors; Cell membrane; Cell physiology; Cell surface; Cells; Code; Data; Disease; Embryo; Endoplasmic Reticulum; Family; Foundations; G-Protein-Coupled Receptors; Glioma; Golgi Apparatus; Guanosine Triphosphate Phosphohydrolases; Hormones; Human; Kidney; Leucine; Mediating; Membrane Proteins; Molecular; Nephrogenic Diabetes Insipidus; Neuroblastoma; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phenylalanine; Positioning Attribute; Regulation; Role; Route; Signal Transduction; Structure; Structure-Activity Relationship; Transport Vesicles; adrenergic; glycosylation; human CCR10 protein; human disease; novel; novel therapeutics; receptor; receptor function; response; trafficking

DESCRIPTION (provided by applicant): G protein-coupled receptors (GPCRs) constitute a super-family of cell surface receptors that regulate a variety of cell functions. Defective export trafficking of GPCRs from the endoplasmic reticulum (ER) through the Golgi to the cell surface is associated with the pathogenesis of a variety of human diseases, nephrogenic diabetes insipidus being one of the best-studied examples. However, the molecular mechanism underlying the export of GPCRs remains poorly understood. Our overall objective is to define the molecular mechanism of GPCR export trafficking and its functional role in regulating cellular responses to hormones and drugs. Under this broad objective, this proposal focuses on two important questions: 1) how GPCRs export from the ER? 2) How GPCRs transport from the ER to the Golgi? Our preliminary data have demonstrated that the motif consisting of a phenylalanine (F) and double leucine (L) spaced by six residues {F(x)6LL} is required for export of the alpha2B-adrenergic (AR) and angiotensin ll-type 1 (AT1R) receptors from the ER. This motif is highly conserved in many GPCRs and thus may provide a common mechanism for their export. Our data also indicate that different GPCRs may use different routes to move to the cell surface in neuroblastoma- glioma NG108 and human embryonic kidney HEK293 cell lines. Non-glycosylated alpha2B-AR uses a novel, as yet undefined pathway. The Specific Aims are: 1) To define the mechanism of the F(x)6LL motif in mediating GPCR export from the ER. We will determine if the F(x)6LL motif is a common code for GPCR export from the ER, determine if the F(x)6LL motif functions as an ER export signal and/or regulates receptor folding, and define the structure-function relationship of the F(x)6LL motif. 2) To define the novel pathway for alpha2B-AR transport from the ER to the Golgi. We will define intracellular compartments and transport vesicles involved in alpha2B-AR transport and determine if glycosylation alters alpha2B-AR transport pathway. These studies will provide new and important information regarding the molecular mechanisms underlying GPCR export and its control on receptor function. Such information may help exploit the possibility of developing new therapeutic strategies for treating disease by targeting GPCR transport.

描述（由申请人提供）：G蛋白偶联受体（GPCR）构成调节多种细胞功能的细胞表面受体超家族。 GPCR 从内质网 (ER) 通过高尔基体到达细胞表面的缺陷输出运输与多种人类疾病的发病机制有关，肾性尿崩症是研究最充分的例子之一。然而，GPCR 输出的分子机制仍然知之甚少。我们的总体目标是确定 GPCR 输出转运的分子机制及其在调节细胞对激素和药物反应中的功能作用。在此总体目标下，该提案重点关注两个重要问题：1）GPCR 如何从 ER 导出？ 2）GPCR如何从内质网运输到高尔基体？我们的初步数据表明，由六个残基 {F(x)6LL} 间隔的苯丙氨酸 (F) 和双亮氨酸 (L) 组成的基序是从 ER 输出 α2B-肾上腺素能 (AR) 和血管紧张素 II-1 型 (AT1R) 受体所必需的。该基序在许多 GPCR 中高度保守，因此可能为其输出提供通用机制。我们的数据还表明，在神经母细胞瘤-神经胶质瘤 NG108 和人胚肾 HEK293 细胞系中，不同的 GPCR 可能使用不同的途径移动到细胞表面。非糖基化 alpha2B-AR 使用一种新颖的、尚未明确的途径。具体目标是： 1) 定义 F(x)6LL 基序介导内质网 GPCR 输出的机制。我们将确定 F(x)6LL 基序是否是 ER 中 GPCR 输出的通用代码，确定 F(x)6LL 基序是否充当 ER 输出信号和/或调节受体折叠，并定义 F(x)6LL 基序的结构功能关系。 2) 定义α2B-AR从内质网转运到高尔基体的新途径。我们将定义参与 alpha2B-AR 运输的细胞内区室和运输囊泡，并确定糖基化是否会改变 alpha2B-AR 运输途径。这些研究将提供有关 GPCR 输出的分子机制及其对受体功能的控制的新的重要信息。这些信息可能有助于开发通过靶向 GPCR 转运来治疗疾病的新治疗策略的可能性。