Cellular Pharmacology of Kappa Opiod Receptor

Kappa 阿片受体的细胞药理学

基本信息

批准号：
7787468
负责人：
LEE-YUAN LIU-CHEN
金额：
$ 29.7万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7787468
关键词：
14-3-3 Proteins ATP phosphohydrolase Abbreviations Absence of pain sensation Adenylate Cyclase Adrenergic Receptor Affect Agonist Amino Acids Anabolism Anti-Anxiety Agents Applications Grants Autophagocytosis Binding Binding Proteins Brefeldin A C-terminal Cell Line Cell Surface Receptors Cell membrane Cell surface Cells Cellular biology Chemicals Chinese Hamster Ovary Cell Coat Protein Complex I Coatomer Protein Cocaine Complementary DNA Coupled Cyclodextrins Diuresis Dominant-Negative Mutation Down-Regulation Dynorphin A Ear Embryo Endocytosis Pathway Endoplasmic Reticulum Endosomes Enhancers Epitopes Equilibrium Event Familiarity Family G Protein-Coupled Receptor Genes G-Protein-Coupled Receptors GTP-Binding Proteins Glutathione S-Transferase Golgi Apparatus Hemagglutinin Horseradish Peroxidase Human Immune response Kidney Light Lipids Lysosomes MAP Kinase Gene Masks Mediating Membrane Microtubule-Associated Proteins Mitogen-Activated Protein Kinases Modification Molecular Chaperones Mus N-ethylmaleimide-sensitive protein Neurotensin Receptors Neurotransmitters Opioid Opioid Receptor Pain Pathway interactions Peptides Pertussis Toxin Pharmaceutical Preparations Pharmacology Phosphoproteins Physiological Play Polyacrylamide Gel Electrophoresis Prostaglandin Receptor Protein Binding Protein Family Proteins Proto-Oncogene Proteins c-raf Quality Control Rattus Recycling Regulation Research Rhodopsin Role Signal Transduction Signaling Molecule Sodium Dodecyl Sulfate-PAGE Sorting - Cell Movement Specificity Stimulus Tail Techniques Testing Transcription Coactivator Transmembrane Domain Visceral Water Yeasts base craving delta opioid receptor dysphoria ezrin glycosylation golgi associated, gamma adaptin homologous, ADP-ribosylation factor interacting protein in vivo insight member moesin mu opioid receptors mutant natural hypothermia novel programs protein complex public health relevance radixin protein receptor receptor binding receptor expression research study response sodium-hydrogen exchanger regulatory factor sortilin trafficking trans-Golgi Network

项目摘要

DESCRIPTION (provided by applicant): The ? opioid receptor (KOPR) is one of the three major types (?, ? and ?) of opioid receptors that mediate effects of opioids in vivo. Activation of KOPR produces analgesia, dysphoria, water diuresis, hypothermia and modulation of immune responses. KOPR antagonists may be potentially useful for curbing cocaine craving and as anti-depressants and anti-anxiety agents. KOPR, a member of the 7TMR family, is coupled through pertussis toxin-sensitive G proteins to a variety of effectors. For 7TMRs the capacity of agonists to modulate downstream signaling molecules depends on the availability of the receptors on cell surface. The number of cell surface 7TMRs reflects a balance between biosynthesis and endocytosis pathways. The post-activation endocytic events have been well-documented; however, regulation along the biosynthesis pathway is much less understood. The focus of this grant application is to characterize the regulation of the KOPR trafficking along the biosynthesis pathway, in particular by the proteins GEC1, sortilin and 14-3-3, which we found to interact with the KOPR and to be involved in KOPR export trafficking. The central hypothesis is that the export trafficking is regulated by molecules interacting with the KOPR. The specific aims are as follows. (1) To delineate mechanisms underlying GEC1-promoted expression and trafficking of the KOPR. The hypothesis that GEC1 enhances KOPR expression by enhancing ATPase activity of NSF, but not by membrane association by lipid conjugation will be tested. (2) To investigate the role of 14-3-3 in the trafficking and cellular pharmacology of the KOPR. The hypothesis is that 14-3-3 proteins bind to KOPR C-terminal domain and/or i3 loop, which masks COPI binding, allowing the KOPR to sort to plasma membranes. (3) To examine the interaction of the KOPR with sortilin and its functional consequences. The hypothesis to be tested is that sortilin binding to the KOPR sorts the receptor to endosomes and lysosomes and this represents a novel post-ER quality control mechanism. We will also examine if the interactions of the KOPR with these proteins affect signaling and regulation. The proposed studies will provide better understanding of cell biology of the KOPR and provide mechanistic insights into regulation of export of the KOPR. In addition, such understanding will have important implications for other membrane bound receptors since these regulatory mechanisms of export are likely to be applicable to some other proteins. PUBLIC HEALTH RELEVANCE: The proposed research will provide better understanding on how cell surface receptor numbers are regulated, which play important roles in response to external stimuli, including drugs and neurotransmitters.

描述（由申请人提供）：？阿片受体（KOPR）是在体内介导阿片类药物作用的三种主要类型（？、？和？）阿片受体之一。 KOPR 的激活会产生镇痛、烦躁、利尿、体温过低和免疫反应调节等作用。 KOPR 拮抗剂可能可用于抑制对可卡因的渴望以及作为抗抑郁药和抗焦虑药。 KOPR 是 7TMR 家族的成员，通过百日咳毒素敏感 G 蛋白与多种效应子偶联。对于 7TMR，激动剂调节下游信号分子的能力取决于细胞表面受体的可用性。细胞表面 7TMR 的数量反映了生物合成和内吞途径之间的平衡。激活后内吞事件已被充分记录；然而，人们对生物合成途径的调控知之甚少。本次拨款申请的重点是表征 KOPR 沿着生物合成途径的运输调节，特别是 GEC1、sortilin 和 14-3-3 蛋白，我们发现它们与 KOPR 相互作用并参与 KOPR 输出贩运。核心假设是出口贩运受到与 KOPR 相互作用的分子的调节。具体目标如下。 (1) 描述 GEC1 促进 KOPR 表达和运输的机制。 GEC1 通过增强 NSF 的 ATP 酶活性而不是通过脂质缀合的膜缔合来增强 KOPR 表达的假设将得到检验。 (2) 研究14-3-3在KOPR的运输和细胞药理学中的作用。假设是 14-3-3 蛋白与 KOPR C 端结构域和/或 i3 环结合，从而掩盖 COPI 结合，从而允许 KOPR 分选至质膜。 (3) 研究 KOPR 与 sortilin 的相互作用及其功能后果。要测试的假设是分拣蛋白与 KOPR 结合将受体分选到内体和溶酶体，这代表了一种新的 ER 后质量控制机制。我们还将检查 KOPR 与这些蛋白质的相互作用是否影响信号传导和调节。拟议的研究将有助于更好地了解 KOPR 的细胞生物学，并为 KOPR 输出的调控提供机制见解。此外，这种理解将对其他膜结合受体产生重要影响，因为这些输出调节机制可能适用于一些其他蛋白质。公共健康相关性：拟议的研究将更好地了解细胞表面受体数量的调节方式，细胞表面受体数量在响应外部刺激（包括药物和神经递质）方面发挥着重要作用。