权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Towards molecular mechanisms of invertebrate Gustatory Receptors

无脊椎动物味觉受体的分子机制

基本信息

批准号：
10064623
负责人：
RACHELLE GAUDET
金额：
$ 19.82万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10064623
关键词：
Behavior Binding Biochemical Biochemistry Blood Buffers Carbon Cells Chemicals Collaborations Complex Cryoelectron Microscopy Crystallography Cues Culicidae Data Dengue Diet Disease Disease Vectors Drosophila melanogaster Family Family member Fructose Future Goals Homo Human Insecta Invertebrates Ion Channel Gating Ligands Malaria Membrane Membrane Proteins Modality Molecular Nramp protein Nutrient Orthologous Gene Physiological Play Positioning Attribute Preparation Property Proteins Protocols documentation Research Personnel Role Sensory Sensory Receptors Sequence Analysis Smell Perception Stimulus Structure System Taste Perception Testing Ticks Variant West Nile virus Work ZIKA chikungunya comparative disorder control extreme temperature feeding fly human disease insect disease vector member olfactory receptor receptor receptor function receptor structure function sensor success sugar vector control

项目摘要

PROJECT SUMMARY Invertebrate Gustatory Receptors (GRs) are a large and evolutionarily diverse family of sensory receptors known to play important roles in invertebrate taste, smell and thermotransduction. Given the importance of these sensory modalities in host-seeking behavior in important humand disease vectors like mosquitoes, GR family members serve as potentially powerful targets for vector control agents. However, little is known about GR structure and function. We propose a physiological and biochemical analysis of members of two GR subfamilies: Gr43a and Gr28bD. These initial studies will serve as a precursor for a subsequent R01 to carry out structural and functional analyses of these GRs. We propose to achieve these goals in two aims: Aim #1: Identify and physiologically characterize multiple orthologs of Gr43a and Gr28bD. Unlike most GRs, Gr43a and Gr28bD orthologs can be functionally characterized in heterologous cells. In aim 1.a., we will express orthologs of these GRs from additional insect species, including disease vectors and extremophiles, in heterologous cells and characterize their physiological properties. This will enable a comparative analysis of sequence and function among each receptor class. Aim #2: Biochemically characterize multiple Gr43a and Gr28bD orthologs. We find Gr43a and Gr28bD orthologs can be partially purified from heterologous cells. In aim 2, we will expand this approach to incorporate additional orthologs characterized in aim 1 and optimize our purification protocol and explore key properties including oligomeric state and thermal stability in various membrane mimics. This will provide important biochemical information about GR complexes and identify orthologs best suited for subsequent structural analysis. The physiological characterization of multiple Gr43a and Gr28bD orthologs will enable direct examination of evolutionary variation and conservation in GR family function. The expression and purification of multiple family members will provide multiple candidates for biochemistry and structural determination, maximizing the likelihood of success of subsequent GR structural determinations.

项目概要无脊椎动物味觉感受器 (GR) 是一个庞大且进化多样的感觉感受器家族已知在无脊椎动物的味觉、嗅觉和热传导中发挥重要作用。鉴于重要性这些感官模式在重要的人类疾病媒介（如蚊子、GR）的宿主寻找行为中家庭成员是病媒控制剂潜在的强大目标。然而，人们对此知之甚少 GR结构和功能。我们建议对两个 GR 成员进行生理生化分析亚科：Gr43a 和 Gr28bD。这些初步研究将作为后续 R01 的前身对这些遗传资源进行结构和功能分析。我们建议通过两个目标来实现这些目标：目标#1：鉴定 Gr43a 和 Gr28bD 的多个直向同源物并对其进行生理学表征。与大多数 GR 不同，Gr43a 和 Gr28bD 直向同源物可以在异源细胞中进行功能表征。瞄准目标 1.a.，我们将表达来自其他昆虫物种的这些遗传资源的直向同源物，包括疾病媒介和异源细胞中的极端微生物并表征其生理特性。这将使每个受体类别之间的序列和功能的比较分析。目标#2：对多个 Gr43a 和 Gr28bD 直系同源物进行生化表征。我们找到 Gr43a 和 Gr28bD 直向同源物可以从异源细胞中部分纯化。在目标 2 中，我们将把这种方法扩展到纳入目标 1 中表征的其他直向同源物并优化我们的纯化方案并探索关键特性，包括各种膜模拟物的寡聚状态和热稳定性。这将提供有关 GR 复合物的重要生化信息，并确定最适合后续研究的直系同源物结构分析。多个 Gr43a 和 Gr28bD 直向同源物的生理特征将能够直接检查 GR家族功能的进化变异和保守。多种蛋白的表达和纯化家庭成员将为生物化学和结构测定提供多个候选者，最大化随后 GR 结构确定成功的可能性。