The Role of Odorant Binding Proteins in Anopheline Mosquito Odor Perception

这里提出的工作将调查的特异性气味识别的疟疾蚊子冈比亚按蚊和相关的气味刺激与可量化的生理和行为反应。这项工作将整合互补的专业知识，在各个相关领域的共同目标，彻底了解化学传感系统和化学传感介导的行为。冈比亚。气味识别特异性很可能编码在气味结合蛋白（OBP）和气味受体（OR）中。最近的证据表明，气味识别是一个协调的过程，需要OBP和OR的组合特异性。OBP是嗅觉系统中第一个遇到气味分子的蛋白质，我们和其他实验室的数据表明，OBP对A.冈比亚。这使得气味结合蛋白成为操纵气味感知的优选靶标。因此，体外表达的OBP将用于高通量筛选测定，以鉴定天然配体并推断已知和新鉴定的配体的结合能力。计算机建模研究使我们能够设计基于诱变的实验来定义与OBP结合口袋相关的氨基酸，并预测具有改进性质的修饰配体。所有体外研究的体内验证将通过电生理和行为测定获得，所述测定采用表达正常水平的OBP或由于RNAi介导的基因表达敲低而缺乏特定OBP的雌性蚊子。从这一多学科方法中获得的知识将有助于确定可干扰雌蚊寻找宿主行为的化学品，从而减少人类宿主与寄生虫病媒之间的接触。

The work proposed here will investigate the specificity of odorant recognition in the malaria mosquito Anopheles gambiae and correlate odor stimuli with quantifiable physiological and behavioral responses. This work will integrate complementary expertise in various relevant fields toward the common goal of thoroughly understanding the chemosensory system and chemosensory-mediated behavior of A. gambiae. Odor recognition specificity is very likely encoded in both odorant binding proteins (OBPs) and odorant receptors (ORs). Recent evidence suggests that odor recognition is a coordinated process that requires the combined specificities contributed by OBPs and ORs. OBPs are the first proteins of the olfactory system to encounter odor molecules, and data from our and other laboratories demonstrate that OBPs are critical for the olfactory response in A. gambiae. This makes the odorant binding proteins a preferred target for manipulating odor perception. Thus, in vitro expressed OBPs will be used in high-throughput screening assays to identify natural ligands and deduce binding capacities for known and newly identified ligands. Computer modeling studies allow us to design mutagenesis-based experiments to define amino acids associated with the OBP binding pocket and predict modified ligands with improved properties. In vivo validation of all in vitro studies will be obtained through electrophysiological and behavioral assays employing female mosquitoes that express normal levels of OBPs or are deficient in a specific OBP as a result of RNAimediated gene expression knockdown. Knowledge gained from this multi-disciplinary approach will lead to the identification of chemicals that can interfere with host seeking behavior by female mosquitoes, reducing the incidence of contact between the human host and the parasite vector.