MOLECULAR AND FUNCTIONAL ANALYSIS OF ATP RECEPTORS

ATP 受体的分子和功能分析

基本信息

批准号：
6639507
负责人：
MARK M VOIGT
金额：
$ 29.6万
依托单位：
SAINT LOUIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-09-15 至 2005-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6639507
关键词：
adenosine triphosphate crosslink density gradient ultracentrifugation gel filtration chromatography immunoprecipitation molecular assembly /self assembly neural transmission neurotransmitter receptor protein structure function purinergic receptor receptor binding receptor expression recombinant proteins site directed mutagenesis tissue /cell culture

项目摘要

Evidence accumulated over the past three decades supports the role of adenosine-tripphosphate (ATP) as a neurotransmitter/neuromodulator in many adrenergic, cholinergic and non-adrenergic/non/cholinergic pathways. Its role as a chemical mediator of intercellular communication has been established in tissues such as the central and peripheral nervous systems, endocrine glands, heart and smooth muscle. However, the mechanisms which mediate ATP's role as a chemical signal is poorly understood. A primary reason for this is the presence of multiple receptor subtypes in most tissues coupled with a lack of subtype-selective agonists and antagonists. Additionally, in some tissues there are high levels of cell-surface enzymes (ecto-ATPases) that can quickly degrade many of the ATP analogues used for receptor classification, resulting in further complications. In order to clearly understand the role of ATP-mediated transmission, it is essential to define the pharmacological and physiological properties of ATP receptors. Currently, ATP receptors fall into one of two classifications, P2Y and P2X receptors. The focus of this grant are the ATP receptors which belong to the P2X category. These receptors are ligand-gated channels that are permeable not only to monovalent cations such as Na+ and K+, but also to Ca2+. In this project, we will circumvent the difficulties associated with investigating multiple receptor subtypes in their native settings through the use of recombinant receptor expression in mammalian cells. This approach allows for the expression of individual receptor subtypes in a controlled environment, thus permitting detailed characterization of the basic properties of a receptor subunit. We have embarked on the investigation of P2X receptors at the molecular and functional level using a compare and contrast methodology. In this study we will elucidate the stoichiometry of receptor assemblies, elucidate the domains involved in subunit interactions and identify amino acid residues that participate in agonist (binding and gating of the channel. These experiments will provide essential information regarding P2X receptor function and the role of ATP as a neurotransmitter in the central and peripheral nervous systems. The overall goal of this project is to provide a clearer understanding of the pharmacological and physiological properties associated with specific P2X receptor subtypes.

在过去的三十年中积累的证据支持腺苷 - 三磷酸盐（ATP）作为神经递质/神经调节剂在许多肾上腺素能，胆碱能/非肾上腺素能/非/非/胆碱能途径中的作用。它作为细胞间交流的化学介质的作用已在中央和周围神经系统，内分泌腺，心脏和平滑肌等组织中建立。但是，介导ATP作为化学信号的作用的机制知之甚少。造成这种情况的主要原因是大多数组织中存在多个受体亚型，以及缺乏亚型选择性激动剂和拮抗剂。此外，在某些组织中，有高水平的细胞表面酶（ECTO-ATPases）可以迅速降解许多用于受体分类的ATP类似物，从而导致进一步的并发症。为了清楚地了解ATP介导的传播的作用，定义ATP受体的药理和生理特性至关重要。目前，ATP受体属于P2Y和P2X受体的两个分类之一。该赠款的重点是属于P2X类别的ATP受体。这些受体是配体门控通道，不仅可以渗透到诸如Na+和K+等单价阳离子，还可以渗透到Ca2+。在该项目中，我们将通过在哺乳动物细胞中使用重组受体表达来研究与在本机环境中研究多个受体亚型相关的困难。这种方法允许在受控环境中表达单个受体亚型，从而允许对受体亚基的基本特性的详细表征。我们已经使用比较和对比方法开始研究P2X受体在分子和功能水平上的研究。在这项研究中，我们将阐明受体组件的化学计量法，阐明与亚基相互作用有关的结构域并确定参与激动剂的氨基酸残基（这些实验的结合和剖道。这些实验将提供有关P2X受体功能的基本信息，以及ATP作为ATP的作用以及在中心和PERIPS中的神经系统中的作用，以使其在中心和Perip sermim cleans cleans cliep ins Provistion clemains clemant clemiac cliep to Projection IS clemariac cliep to nection in Cleare clemiac clemain clemant IS肯定的目标。与特定P2X受体亚型相关的生理特性。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Co-expression of P2X1 and P2X5 receptor subunits reveals a novel ATP-gated ion channel.

P2X1 和 P2X5 受体亚基的共表达揭示了一种新型 ATP 门控离子通道。

DOI：
10.1124/mol.54.6.989
发表时间：
1998
期刊：
Molecular pharmacology
影响因子：
3.6
作者：
Torres,GE;Haines,WR;Egan,TM;Voigt,MM
通讯作者：
Voigt,MM

Properties of the novel ATP-gated ionotropic receptor composed of the P2X(1) and P2X(5) isoforms.

DOI：
发表时间：
1999-10
期刊：
Molecular pharmacology
影响因子：
3.6
作者：
W. Haines;G. Torres;M. Voigt;Terrance M. Egan
通讯作者：
W. Haines;G. Torres;M. Voigt;Terrance M. Egan

Molecular cloning and functional characterization of the zebrafish ATP-gated ionotropic receptor P2X(3) subunit.

斑马鱼 ATP 门控离子受体 P2X(3) 亚基的分子克隆和功能表征。

DOI：
10.1016/s0014-5793(00)01685-9
发表时间：
2000
期刊：
FEBS letters
影响因子：
3.5
作者：
Egan,TM;Cox,JA;Voigt,MM
通讯作者：
Voigt,MM

A domain contributing to the ion channel of ATP-gated P2X2 receptors identified by the substituted cysteine accessibility method.

通过取代半胱氨酸可及性方法鉴定出对 ATP 门控 P2X2 受体离子通道有贡献的结构域。

DOI：
10.1523/jneurosci.18-07-02350.1998
发表时间：
1998
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Egan,TM;Haines,WR;Voigt,MM
通讯作者：
Voigt,MM

Topological analysis of the ATP-gated ionotropic [correction of ionotrophic] P2X2 receptor subunit.

ATP 门控离子型 [离子型校正] P2X2 受体亚基的拓扑分析。