Conformations and interactions of insect olfactory receptor proteins

描述(申请人提供)：虫媒疾病是一个重大的世界性健康问题，占全球传染病总负担的近20%。这些疾病中的许多都有可能传播到美国。由于昆虫依靠化学感官来定位、取食和交配，因此了解昆虫嗅觉的分子基础将有助于对昆虫病媒的成功攻击。至少有四种独立的蛋白质参与昆虫信息素的检测：i)信息素结合蛋白；ii)气味特异的嗅觉受体(OR)；iii)在大多数嗅觉感受器神经元中表达的co-OR；以及iv)CD36清道夫受体家族的成员-感觉神经元膜蛋白(SNMP)。信息素结合蛋白为水溶性蛋白，OR、co-OR和SNMP3种为膜蛋白。各种证据支持这样的假设，即昆虫信息素反应是这四种蛋白质成分相互作用的结果。然而，完整的膜蛋白组分的三维结构尚不清楚，也不知道这些蛋白在嗅觉信号接收过程中是否发生了构象变化。此外，关于水溶性和膜结合组分之间相互作用的性质的信息也很少。在拟议的实验中，发光共振能量转移(LRET)将被用来测量果蝇黑腹果蝇中参与信息素接收的蛋白质上指定位置之间的距离。这些位点将被基因插入到信息素特异性受体Or67d、辅助受体Or83d、SNMP和信息素结合蛋白LUSH中。在Sf9细胞中表达后，发光测量将提供关于受体蛋白的空间排列以及在信息素接收过程中是否发生构象变化的信息。这一结果将有助于为未来开发通过操纵嗅觉系统来控制昆虫媒介的方法提供基础。公共卫生相关性：虫媒疾病是一个主要的世界性卫生问题，占全世界传染病总负担的近20%，其中许多疾病有可能传播到美国。由于昆虫依靠化学感官来定位、取食和交配，因此了解昆虫嗅觉的分子基础将有助于对昆虫病媒的成功攻击。此外，昆虫信息素受体的一个组成部分与人类蛋白质相似，涉及动脉粥样硬化、炎症反应、代谢综合征、心肌病、高频听力损失和高谢病。

DESCRIPTION (provided by applicant): Insect-borne disease is a major world health problem, accounting for nearly 20% of the total world- wide infectious disease burden. Many of these diseases are capable of spreading to the United States. Because insects depend on chemical senses for host location, feeding, and mating, a successful assault on insect disease vectors will benefit from understanding the molecular basis of insect olfaction. At least four separate proteins appear to be involved in insect pheromone detection: i) a pheromone-binding protein; ii) an odor-specific olfactory receptor (OR); iii) a co-OR that is expressed in most olfactory receptor neurons; and iv) sensory neuron membrane protein (SNMP), a member of the CD36 scavenger receptor family. Pheromone-binding proteins are water-soluble proteins; and OR, co-OR and SNMP are integral membrane proteins. Various lines of evidence support the hypothesis that the insect pheromone response results from the interaction of these four protein components. However, the three-dimensional structures of the integral membrane protein components are unknown, and it is not known whether these proteins undergo conformational changes during reception of olfactory signals. Also, little information is known about the nature of the interaction between the water-soluble and membrane-bound components. In the proposed experiments, luminescence resonance energy transfer (LRET) will be used to measure distances between defined sites on proteins involved in pheromone reception in the fruit fly, Drosophila melanogaster. The sites will be genetically inserted into the pheromone-specific receptor Or67d, the co-receptor Or83d, SNMP, and the pheromone-binding protein LUSH. After expression in Sf9 cells, luminescence measurements will provide information about the spatial arrangement of the receptor proteins and whether any conformational changes occur during pheromone reception. The results will help provide the basis for future development of methods to control insect vectors through manipulation of the olfactory system. PUBLIC HEALTH RELEVANCE: Insect-borne disease is a major world health problem, accounting for nearly 20% of the total world- wide infectious disease burden, with many of these diseases capable of spreading to the United States. Because insects depend on chemical senses for host location, feeding, and mating, a successful assault on insect disease vectors will benefit from understanding the molecular basis of insect olfaction. Also, one component of the insect pheromone receptor is similar to human proteins involved in atherosclerosis, inflammatory response, metabolic syndrome, cardiomyopathies, high frequency hearing loss, and Gaucher's disease.