The Role of Perireceptive Mucoidal Proteins in Defining Vomeronasal Organ Function in a Macrosmatic Mammal

周围感受粘液蛋白在定义大型哺乳动物犁鼻器官功能中的作用

• 批准号: 0544191
• 负责人: L.E.L. Rasmussen
• 金额: $ 38.83万
• 依托单位: Oregon Health & Science University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0544191&HistoricalAwards=false
• 关键词: Role; Perireceptive; Mucoidal; Proteins; Defining

AWARD ABSTRACTMounting evidence exists for proteins being central to pheromone transport. However, an unresolved question in mammalian olfaction is how these biologically relevant odorants traverse perireceptive olfactory mucous environments to interact with cognate odorant receptors? The extensive dual olfactory system of the macrosmatic Asian elephant (Elephas maximus) offers novel and valid investigative advantages to resolve the source-to-sink molecular trail of two identified pheromones: Z-7-dodecenyl acetate (Z7-12:Ac) and frontalin. The recent discovery of two natural enantiomeric forms of frontalin required in a specific ratio for biological activity is a significant mammalian finding, providing an exquisitely precise chemical model. Our productive international collaboration has described the functional roles of an albumin transporter and an odorant-binding protein sequestrant prior to the final step when the pheromone transits through the long vomeronasal duct before contacting vomeronasal organ (VNO) receptors. Preliminary results support the hypothesis of passage facilitation by a mucous protein.Three research questions are explored: (1) Is there a particular protein in VNO ductal mucus that specifically binds Z7-12:Ac and frontalin enantiomers, consequently transporting them? The study will employ plentiful synthetic pheromones and innovative radiolabelled and photoaffinity visualization analogs. (2) What is the identity of this vomeronasal ductal protein (VDP) and does its recombinant form retain bioactivity? After the protein has been isolated and identified and its gene cloned and sequenced, sufficient amounts will be expressed recombinantly. (3) Does the VDP with its ligand have appropriate biochemical properties (e.g., binding kinetics) and bioactive attributes to be the VNO ductal transporter?In elephants, spatial and temporal slowing and separation of individual biochemical events allows entire sequences of perireceptive events to be discerned as discrete steps. In vitro kinetic analyses of VDP-pheromone complexes will be conducted in parallel with in vivo monitoring of discrete-in-time chemosensory responses and behaviors.This study is multidisciplinary and integrates diverse scientific disciplines, from molecular biology and proteomics through chemosensory bioassays and behavior, integrating the common theme of biochemistry, providing a unique opportunity to quantify responses to pheromones, and elucidating specific and functional protein-ligand associations and transport phenomena.Investigations by the international team of Drs. Greenwood and Rasmussen will continue the dual tradition of broadly impacting science and practically helping conservation. Advances in knowledge of olfaction in the endangered Asian elephant have proximate utility in practical deterrent development, thereby reducing human-elephant conflict. Ultimately, how binding proteins interact with pheromones at the level of chirality will contribute fundamental insights into vertebrate olfaction. In the broadest sense, this research will benefit human society by helping to preserve an extraordinary species that has interacted with man for thousands of years.

奖项摘要 越来越多的证据表明蛋白质是信息素运输的核心。 然而，哺乳动物嗅觉中一个未解决的问题是这些生物学相关的气味剂如何穿过嗅觉粘液环境与同源气味受体相互作用？ 亚洲象 (Elephas maximus) 广泛的双嗅觉系统为解析两种已识别的信息素：Z-7-十二碳烯基乙酸酯 (Z7-12:Ac) 和额叶素的从源到库的分子轨迹提供了新颖且有效的研究优势。 最近发现了两种天然对映体形式的额叶蛋白，其生物活性需要特定的比例，这是哺乳动物的一项重大发现，提供了一个极其精确的化学模型。 我们富有成效的国际合作描述了白蛋白转运蛋白和气味结合蛋白螯合剂在最后一步之前的功能作用，即信息素在接触犁鼻器 (VNO) 受体之前穿过长犁鼻管。 初步结果支持粘液蛋白促进通过的假说。探讨了三个研究问题：(1) VNO 导管粘液中是否存在一种特定的蛋白质，可以特异性结合 Z7-12:Ac 和额叶蛋白对映体，从而运输它们？ 该研究将采用大量的合成信息素和创新的放射性标记和光亲和可视化类似物。 (2) 这种犁鼻导管蛋白 (VDP) 的身份是什么？其重组形式是否保留生物活性？ 在蛋白质被分离和鉴定以及其基因被克隆和测序之后，将重组表达足够量的蛋白质。 (3) VDP 及其配体是否具有适当的生化特性（例如，结合动力学）和生物活性属性，​​可以成为 VNO 导管转运蛋白？在大象中，空间和时间的减慢以及单个生化事件的分离允许将整个感知事件序列识别为离散步骤。 VDP-信息素复合物的体外动力学分析将与体内离散时间化学感应反应和行为的监测同时进行。这项研究是多学科的，整合了不同的科学学科，从分子生物学和蛋白质组学到化学感应生物测定和行为，整合了生物化学的共同主题，为量化化学感应反应和行为提供了独特的机会。 信息素，并阐明特定的功能性蛋白质配体关联和运输现象。格林伍德和拉斯穆森将延续广泛影响科学和切实帮助保护的双重传统。 濒临灭绝的亚洲象嗅觉知识的进步对于实际威慑发展具有直接效用，从而减少人象冲突。 最终，结合蛋白如何在手性水平上与信息素相互作用将有助于对脊椎动物嗅觉的基本了解。 从最广泛的意义上讲，这项研究将通过帮助保护与人类互动了数千年的非凡物种而造福人类社会。