Elucidating the structural determinants of odor specificity in insect olfactory receptors

阐明昆虫嗅觉受体气味特异性的结构决定因素

• 批准号: 10611096
• 负责人: Josefina Inés Del Marmol
• 金额: $ 24.9万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-05 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611096
• 关键词: Address; Advisory Committees; Aedes; Affect; Alleles; Amino Acid Receptors; Amino Acid Sequence; Animals; Appointment; Behavioral Assay; Binding; Biochemical; Biological Assay; Biophysics; Chemicals; Complement; Complex; Computer Models; Cryoelectron Microscopy; Culicidae; Data; Dengue Fever; Detection; Development; Development Plans; Discrimination; Disease; Drosophila genus; Drosophila melanogaster; Electrophysiology (science); Elements; Engineering; Environment; Exhibits; Family; Foundations; Functional Imaging; Genetic; Genetic Polymorphism; Goals; Human; Impairment; In Vitro; Insect Repellents; Insecta; Leadership; Ligand Binding; Ligands; Light; Link; Logic; Malaria; Mammals; Manuscripts; Mentors; Mentorship; Modality; Molecular; Mutagenesis; Mutation; Neurons; Odorant Receptors; Odors; Olfactory Pathways; Persons; Phase; Positioning Attribute; Preparation; Property; Research; Research Project Grants; Resolution; Resources; Ruta; Sensory; Smell Perception; Specificity; Structure; System; Testing; Training; Universities; Variant; Work; Yellow Fever; career development; combinatorial; drug discovery; flexibility; in vivo; insect disease vector; insight; molecular modeling; novel; olfactory receptor; olfactory stimulus; preference; prevent; programs; rational design; receptor; receptor binding; response; skills; small molecule; structural biology; tool; zika fever

Project Summary The sense of smell provides animals with vital information about their environment. At the molecular level, olfactory stimuli consist of thousands of distinct small molecules that share no common chemical feature other than being volatile. To contend with this diversity, insects and mammals alike have evolved large families of olfactory receptors (ORs) that operate in a combinatorial way, whereby some receptors are broadly activated by many different odorants and others are exquisitely tuned to a specific compound. The molecular mechanisms that endow ORs with such diverse ligand-binding properties remain unknown, largely because the isolation and structural characterization of ORs has been a decades-long technological challenge. Using cryo-electron microscopy, I recently determined the first atomic-resolution structure of an insect OR bound to an odor. By determining the structures of additional insect ORs with broad and specific ligand-binding properties, the proposed research project will elucidate the molecular mechanism of odor detection and discrimination. To this end, the K99 mentored phase (Aim 1) will reveal the atomic structures of two primitive insect ORs with different ligand specificities unbound and in complex with multiple odor ligands, shedding light on the molecular determinants that underlie broad or specific odor selectivity. Next, the R00 independent phase (Aim 2) will focus on ORs from disease-carrying mosquitoes that participate in human host-seeking. These ORs exhibit small polymorphisms that drastically affect their ability to detect human odors. Elucidation of the atomic structure of these mosquito ORs bound to human volatiles will illuminate the molecular properties that enable mosquitoes to detect and prey on humans. Together, this research program will lend fundamental insight into the normal function of sensory processing of olfactory information. Additionally, as insect ORs are critically involved in human host-seeking that facilitates the spread of insect-borne diseases, this work will provide a foundation for the development of novel insect repellents that could curb diseases such as malaria, Zika, and dengue fever. The proposed development plan complements my training in structural biology and biophysics with computational modeling of receptor-ligand interactions and in vivo assays in Drosophila. At the end of the mentored phase, I will be equipped with the necessary tools to conduct comprehensive structural and functional studies of odor detection and discrimination by insect vectors of disease. To achieve these goals, I will take advantage of the extensive resources of the Rockefeller University, the mentorship of Dr. Vanessa Ruta and the appointment of an Advisory Committee that will lend expertise in key aspects of the project and career development. Additionally, with the support and resources from the MOSAIC/UE5 network I will expand my mentorship and leadership skills to successfully transition to an independent position while continuing my ongoing efforts to enhance diversity in the biomedical workforce.

项目概要 嗅觉为动物提供有关其环境的重要信息。在分子水平上， 嗅觉刺激由数千个不同的小分子组成，这些小分子没有共同的化学特征 而不是不稳定。为了应对这种多样性，昆虫和哺乳动物都进化出了大家族 嗅觉受体（OR）以组合方式运作，其中一些受体被广泛激活 许多不同的气味剂和其他气味剂都经过精心调整以适应特定的化合物。分子机制 赋予 OR 如此多样化的配体结合特性仍然未知，很大程度上是因为分离和 OR 的结构表征一直是长达数十年的技术挑战。使用低温电子 通过显微镜，我最近确定了昆虫或与气味结合的第一个原子分辨率结构。经过 确定具有广泛和特异性配体结合特性的其他昆虫 OR 的结构， 拟议的研究项目将阐明气味检测和辨别的分子机制。 为此，K99 指导阶段（目标 1）将揭示两种原始昆虫 OR 的原子结构 不同的配体特异性未结合并与多种气味配体复合，揭示了分子 广泛或特定气味选择性的决定因素。接下来，R00独立阶段（目标2）将重点关注 参与人类寻找宿主的携带疾病的蚊子的OR。这些 OR 表现出较小的 多态性极大地影响了它们检测人类气味的能力。原子结构的阐明 这些与人类挥发物结合的蚊子 OR 将阐明使蚊子能够 发现并捕食人类。总之，该研究计划将为正常情况提供基本见解 嗅觉信息的感觉处理功能。此外，由于昆虫 OR 关键参与 寻找人类宿主促进了虫媒疾病的传播，这项工作将为 开发可以遏制疟疾、寨卡病毒和登革热等疾病的新型驱虫剂。 拟议的发展计划补充了我在结构生物学和生物物理学方面的培训 果蝇受体-配体相互作用的计算模型和体内测定。结束时 指导阶段，我将配备必要的工具来进行全面的结构和功能 疾病昆虫媒介的气味检测和辨别研究。为了实现这些目标，我将采取 得益于洛克菲勒大学的丰富资源、Vanessa Ruta 博士的指导和 任命一个咨询委员会，该委员会将提供项目和职业关键方面的专业知识 发展。此外，借助 MOSAIC/UE5 网络的支持和资源，我将扩展我的 指导和领导技能，以成功过渡到独立职位，同时继续我的工作 不断努力增强生物医学劳动力的多样性。