Proteomic Characterization of the Sigma-1 Receptor and Its Signaling Complex

Sigma-1 受体及其信号复合物的蛋白质组学表征

基本信息

批准号：
7846795
负责人：
CARTHENE R BAZEMORE-WALKER
金额：
$ 20.24万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7846795
关键词：
Acute Affinity Amino Acid Sequence Antisense Oligonucleotides Arts Behavior Binding Biological Biological Process Brain Brain Stem Cell membrane Cocaine Collaborations Complex Comprehension Cytoskeletal Proteins DNA Sequence Rearrangement Data Drug Addiction Drug abuse Environment Event Family Funding Goals Health High Pressure Liquid Chromatography Human Ion Channel Knowledge Label Lead Learning Ligand Binding Ligands Lipids Literature Locomotion Mass Spectrum Analysis Membrane Memory Methamphetamine Methods Mission Molecular Moods Movement Neuraxis Neurology Neurons Neurotransmitter Receptor Neurotransmitters Opioid Receptor Outcomes Research Pathway interactions Peripheral Pharmacology Physiological Play Post-Translational Protein Processing Posture Process Protein Isoforms Protein Sequence Analysis Proteins Proteomics Public Health Publishing Receptor Signaling Regulation Research Research Activity Research Personnel Resource Sharing Resources Role Scheme Science Signal Pathway Signal Transduction Structure Substance abuse problem System Testing Therapeutic Intervention Tissues Translating Walkers addiction base drug induced behavior drug of abuse human RIPK1 protein improved innovation instrumentation novel protein protein interaction psychostimulant public health relevance receptor receptor binding receptor function sigma receptors sigma-1 receptor sigma-2 receptor small molecule therapeutic target

项目摘要

DESCRIPTION (provided by applicant): A fundamental understanding of how the sigma-1 receptor is regulated at the protein level, and how this impacts its function, is still missing. This is important because the sigma receptors represent a novel class of proteins that when activated by psychostimulants, such as cocaine and methamphetamine, contribute to both short- and long-term adaptations in the brain. Our long-term goal is to elucidate the regulatory mechanisms controlling activation of sigma receptors as a prerequisite to fully understanding sigma receptor function. The overall objective of this R03 application, which is a first step toward attainment of our long-term goal, is to fully characterize the sigma-1 receptor and its "signaling complex." The central hypothesis of this application is that the sigma-1 receptor utilizes a network of interacting proteins to translate ligand binding events into biological action and this signaling network is subject to regulation by post-translational protein modifications and differential protein-protein interactions. The rationale for the proposed research is that it will be possible to study sigma-1 receptor regulation of additive behaviors more thoroughly once we understand how its signaling is controlled. Since the sigma-1 receptor is associated with various biological processes, this knowledge will potentially lead to successful strategies for targeting structurally (and functionally) diverse isoforms specifically. The proposed research is relevant to NIH's mission because it will expand our basic knowledge and potentially improve the human health. Two specific aims will be pursued in order to test our central hypothesis and accomplish the objective of this application: 1) Identify sigma-1 receptor post-translational modifications in the presence and absence of ligands; and 2) Identify sigma-1 receptor interacting-proteins in the presence and absence of ligands. We will accomplish this by using tailored purification schemes and specific labeling strategies in conjunction with optimized HPLC gradients and state-of-the art mass spectrometry instrumentation. The proposed research is significant because it will lead to a better fundamental comprehension of a novel class of membrane bound receptors and expand our understanding of how the receptor is regulated by drugs of abuse. PUBLIC HEALTH RELEVANCE: Sigma-1 receptors, potentially important therapeutic targets for drug addiction and abuse, will be comprehensively characterized in the proposed studies. The proposed research is relevant to public health because the results are expected to lead to a better understanding of how these proteins can be manipulated to treat substance abuse.

描述（由申请人提供）：仍然缺少对Sigma-1受体如何调节Sigma-1受体的基本理解，以及这如何影响其功能。这很重要，因为Sigma受体代表了一种新型的蛋白质，当通过心理刺激剂（例如可卡因和甲基苯丙胺）激活时，会导致大脑中的短期和长期适应性。我们的长期目标是阐明控制Sigma受体激活的调节机制，作为完全理解Sigma受体功能的先决条件。该R03应用的总体目标是实现我们长期目标的第一步，是完全表征Sigma-1受体及其“信号传导复合物”。该应用的中心假设是Sigma-1受体利用相互作用的蛋白质网络将配体结合事件转化为生物学作用，并且该信号网络受翻译后蛋白质修饰和差异蛋白质蛋白质相互作用的调节。拟议的研究的理由是，一旦我们了解如何控制其信号传导，就可以更彻底地研究添加剂行为的Sigma-1受体调节。由于Sigma-1受体与各种生物学过程有关，因此该知识可能会导致成功靶向结构（功能和功能）多样化的同工型的成功策略。拟议的研究与NIH的使命有关，因为它将扩大我们的基本知识并有可能改善人类健康。为了检验我们的中心假设并实现了本应用的目标：1）在存在和不存在配体的情况下，将识别Sigma-1受体后翻译后修饰； 2）在存在和不存在配体的情况下鉴定Sigma-1受体相互作用蛋白。我们将通过使用量身定制的纯化方案和特定的标记策略以及优化的HPLC梯度和最先进的质谱仪器来实现这一目标。拟议的研究很重要，因为它将导致对新型膜结合受体的基本理解，并扩展我们对受体如何受滥用药物调节的理解。公共卫生相关性：在拟议的研究中，将全面表征Sigma-1受体，潜在的药物成瘾和滥用治疗靶标。拟议的研究与公共卫生有关，因为预计结果将使人们更好地了解如何操纵这些蛋白质以治疗药物滥用。