Structure and Function of TRP Thermosensation

TRP 热敏的结构和功能

• 批准号: 7799549
• 负责人: Jorg Grandl
• 金额: $ 5.99万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7799549
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Affect; Amino Acids; Binding; Capsaicin; Cell membrane; Charge; Chemicals; Communities; Data; Disease; Dose; Engineering; Esthesia; Goals; Heating; Ion Channel; Ions; Lead; Learning; Libraries; Ligands; Light; Measures; Menthol; Molecular; Mutate; Mutation; Nature; Nervous system structure; Pain; Phenotype; Point Mutation; Property; Proteins; Publishing; Qualifying; Research; Screening Result; Sensory; Side; Skin Temperature; Structure; TRPV1 gene; Temperature; Temperature Sense; Testing; chronic pain; follow-up; insight; interest; mutant; receptor; research study; response; theories; tool; voltage

DESCRIPTION (provided by applicant): The research goal is to understand how transient receptor potential (TRP) channels sense temperature. In this project, the central hypothesis is that unique residues (domains) within the temperature-activated TRP channels enable these proteins to sense changes in temperature and to gate ion cun-ents through the plasma membrane. Specific functional domains of the temperature-activated ion channels TRPM8 and TRPV1 should be found by usage of large-scale random mutant libraries. Specific aim 1: screen large mutant libraries of the heat- and cold-activated ion-channels TRPV1 and TRPM8 to find mutations that specifically affect temperature-activation, but not chemical activation. This aim will test, if domains exist in these channels that are specifically required for temperature-activation. Specific aim 2: detailed electrophysiological examination of these mutants to verify screening results and to find out if temperature-mutations uncouple the mechanism of voltage-activation. This might shed light on the question, if voltage-activation underlies the mechanism of temperature-activation, as was previously proposed, but is still questioned. Specific aim 3: mutation of identified residues into all other 19 amino-acids to learn what specific molecular interactions are associated with the temperature-deficient phenotype. These experiments could give detailed insight into the molecular mechanism and lead to new, testable theories of the mechanism of temperature-activation. Chronic pain is associated with many different diseases and thermosensation represents a significant component of pain sensation. Therefore, a molecular understanding of thermosensation is important and relevant to the field of pain. I expect this study to significantly advance the understanding of how we sense temperature at the molecular level.

描述（由申请人提供）：研究目标是了解瞬态受体电位（TRP）通道如何感知温度。在该项目中，中心假设是温度激活的TRP通道内的独特残基（域）使这些蛋白质能够感知温度的变化，并通过质膜通过栅极离子CUN含量。温度激活的离子通道的特定功能域TRPM8和TRPV1应通过使用大规模随机突变库来找到。特定目标1：屏幕热激活的离子通道TRPV1和TRPM8的大型突变库，以找到特异性影响温度激活但没有化学激活的突变。如果在这些特定温度激活所需的这些通道中存在域，则该目标将测试。具体目的2：对这些突变体的详细电生理检查，以验证筛选结果，并找出温度突变是否使电压激活的机理失去。这可能会阐明这个问题，如果电压激活是像以前提出的那样，但仍受到质疑，是否是温度激活的机理。特定目的3：已鉴定的残基突变到所有其他19个氨基酸中，以了解哪些特定分子相互作用与温度缺陷型表型相关。这些实验可以详细介绍分子机制，并导致有关温度激活机制的新的，可检验的理论。慢性疼痛与许多不同的疾病有关，热效代表了疼痛感觉的重要组成部分。因此，对热增敏的分子理解与疼痛场很重要，并且与疼痛领域有关。我希望这项研究能够显着提高我们如何在分子水平上感知温度的理解。