Chemical Sensing: Linking sequence, mechanisms and inhibition

化学传感：连接序列、机制和抑制

• 批准号: 1807722
• 负责人: Smita Mohanty
• 金额: $ 63万
• 依托单位: Oklahoma State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1807722&HistoricalAwards=false
• 关键词: Chemical; Sensing; Linking; sequence; mechanisms

Smita Mohanty of Oklahoma State University and Eric Anslyn of the University of Texas, Austin, are supported by an award from the Chemistry of Life Processes Program in the Division of Chemistry to investigate the biochemical and physiological mechanisms of molecular signaling in two lepidopteran insect species (moths). They seek to understand the molecular detail of how an odor is detected and communicated from the female moth to the male moth within a species during the mating process. The ability to respond to chemical stimuli such as odors is a fundamental behavior of all organisms. This project is utilizing structural and biochemical experiments to discover the basic mechanism of odor communication in two moth species, one of which is a voracious pest of crops and stored foods. Computationally-designed molecules are being synthesized and tested in the laboratory for their ability to competitively bind to odor-binding protein molecules to inhibit the sensing of female-secreted scent by the male, and thus disrupt the mating process. Knowledge gained from this research is helping bridge the fundamental gap in knowledge of the chemical and mechanistic details of sense of smell in animals, as well as paving the way for the development of novel, species-specific, and environmentally-friendly odor mimetics as alternatives to harmful pesticides, for insect control. Students at the undergraduate and graduate levels are being trained in state-of-the-art instrumentation, thus preparing the future generation of teachers, researchers, and innovators.Insects use insoluble fatty acid (FA) derivatives as highly-specific signaling molecules. Lepidopteran insects serve as a model for studying the biochemical and physiological mechanisms of FA signaling. Odor/pheromone binding proteins (OBP/PBP) ferry the fatty acid odor to the odorant receptor across the aqueous sensillar lymph that surrounds the dendrites of odor-sensitive olfactory neurons. The details of the mechanism of lipid transfer to the olfactory receptor neuron by OBPs/PBPs remain largely unknown. This project is using the combined tools and techniques of biochemistry, biophysics, molecular biology, chemical modeling, and synthetic organic chemistry to identify the chemical signatures in the protein sequences that dictate the mechanism of lipid binding and release by PBPs in two lepidopteran species: Anthereae polyphemus and Ostrinia furnacalis. The current model for lepidopteran PBPs, including Anthereae polyphemus PBP (ApolPBP1), posits a pH-dependent conformational switch for the uptake and release of the FA signal at the sensory neuron membrane. However, preliminary data for Ostrinia furnacalis PBP (OfurPBP2) suggests that this protein binds lipid and releases it through a distinct, novel mechanism. A strategy of successive mutations in ApolPBP1 residues is being used to systematically transform ApolPBP1 into an OfurPBP2 mimic, and characterize the mutants at each step using a multidisciplinary approach, to gain insight into the unique structure-function relationship and mechanism of lipid binding and release in Ostrinia furnacalis. Nature-inspired design of fatty-acid mimetics is being performed through structure-based organic synthesis, to develop a competitive inhibitor of ApoIPBP1 as a model system, and establish a design strategy for odor inhibition in the Ostrinia furnicalis pest. This combined, complementary data on two key PBP proteins is deepening the understanding of distinct mechanisms of chemical sensing, and providing a foundation for the rational design of eco-friendly biomimetic inhibitors for insect control. Educational and outreach efforts are leveraging the successful NSF REU program at Oklahoma State to recruit and train undergraduates techniques, with students actively recruited from HBCUs (Historically Black Colleges and Universities) and PUIs (Primarily Undergraduate Institutions).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

俄克拉荷马州立大学的斯米塔·莫汉蒂（Smita Mohanty）和德克萨斯大学奥斯汀分校的埃里克·安斯林（Eric Anslyn）获得化学系生命过程化学项目的资助，研究两种鳞翅目昆虫（飞蛾）分子信号传导的生化和生理机制。 他们试图了解一个物种在交配过程中如何检测气味并将其从雌蛾传递给雄蛾的分子细节。对气味等化学刺激做出反应的能力是所有生物体的基本行为。该项目正在利用结构和生化实验来发现两种蛾类的气味传播的基本机制，其中一种是一种贪吃农作物和储存食物的害虫。计算设计的分子正在实验室中合成和测试，以检测它们竞争性地与气味结合蛋白分子结合的能力，以抑制雄性对雌性分泌的气味的感知，从而扰乱交配过程。 从这项研究中获得的知识有助于弥合动物嗅觉化学和机械细节知识的根本差距，并为开发新型、物种特异性和环境友好的气味模拟物作为有害杀虫剂的替代品并用于昆虫控制铺平道路。本科生和研究生级别的学生正在接受最先进仪器的培训，从而为下一代教师、研究人员和创新者做好准备。昆虫使用不溶性脂肪酸 (FA) 衍生物作为高度特异性的信号分子。鳞翅目昆虫是研究 FA 信号传导生化和生理机制的模型。气味/信息素结合蛋白 (OBP/PBP) 将脂肪酸气味穿过气味敏感嗅觉神经元树突周围的水性感觉淋巴传递到气味受体。 OBPs/PBPs 将脂质转移至嗅觉受体神经元的机制细节仍然很大程度上未知。该项目利用生物化学、生物物理学、分子生物学、化学建模和有机合成化学的综合工具和技术来识别蛋白质序列中的化学特征，这些特征决定了两种鳞翅目物种（Anthereae polyphemus 和 Ostrinia fnacalis）中 PBP 脂质结合和释放的机制。当前的鳞翅目 PBP 模型，包括 Anthereae polyphemus PBP (ApolPBP1)，假定感觉神经元膜上 FA 信号的摄取和释放存在 pH 依赖性构象开关。然而，亚洲玉米螟 PBP (OfurPBP2) 的初步数据表明，这种蛋白质结合脂质并通过一种独特的新颖机制将其释放。 ApolPBP1残基的连续突变策略被用于系统地将ApolPBP1转化为OfurPBP2模拟物，并使用多学科方法表征每个步骤的突变体，以深入了解亚洲玉米螟独特的结构-功能关系和脂质结合和释放的机制。 受自然启发的脂肪酸模拟物设计正在通过基于结构的有机合成进行，以开发ApoIPBP1的竞争性抑制剂作为模型系统，并建立亚洲玉米螟害虫气味抑制的设计策略。 这种关于两种关键 PBP 蛋白的综合互补数据正在加深对化学传感不同机制的理解，并为合理设计用于昆虫控制的环保仿生抑制剂奠定了基础。 教育和外展工作正在利用俄克拉荷马州成功的 NSF REU 计划来招募和培训本科生技术，学生积极从 HBCU（历史悠久的黑人学院和大学）和 PUI（主要本科院校）招募。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查进行评估，被认为值得支持 标准。

项目成果

Backbone and side chain NMR assignments and secondary structure calculation of the pheromone binding protein3 of Ostrinia nubilalis, an agricultural pest

• DOI: 10.1007/s12104-023-10145-3
• 发表时间: 2023-07-27
• 期刊: BIOMOLECULAR NMR ASSIGNMENTS
• 影响因子: 0.9
• 作者: Al-Danoon，Omar;Mohanty，Smita
• 通讯作者: Mohanty，Smita

NMR and MD simulations reveal the impact of the V23D mutation on the function of yeast oligosaccharyltransferase subunit Ost4

NMR 和 MD 模拟揭示了 V23D 突变对酵母寡糖转移酶亚基 Ost4 功能的影响

• DOI: 10.1093/glycob/cwab002
• 发表时间: 2021
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: Chaudhary, Bharat P;Zoetewey, David L;McCullagh, Martin J;Mohanty, Smita
• 通讯作者: Mohanty, Smita

1H, 13C, and 15N resonance assignment and secondary structure of the pheromone binding protein from the agricultural pest Ostrinia furnacalis (OfurPBP2)

农业害虫玉米螟信息素结合蛋白 (OfurPBP2) 的 1H、13C 和 15N 共振分配和二级结构

• DOI: 10.1007/s12104-020-09930
• 发表时间: 2020
• 期刊: Biomolecular NMR assignments
• 影响因子: 0.9
• 作者: Dahal, S.

1H, 13C, and 15N resonance assignment and secondary structure of the pheromone-binding protein2 from the agricultural pest Ostrinia furnacalis (OfurPBP2)

• DOI: 10.1007/s12104-020-09930-1
• 发表时间: 2020-04-01
• 期刊: BIOMOLECULAR NMR ASSIGNMENTS
• 影响因子: 0.9
• 作者: Dahal, Salik R.;Lewellen, Jacob L.;Mohanty, Smita
• 通讯作者: Mohanty, Smita

Ostrinia furnacalis PBP2 solution NMR structure: Insight into ligand binding and release mechanisms

• DOI: 10.1002/pro.4438
• 发表时间: 2022-09
• 期刊: Protein Science
• 影响因子: 8
• 作者: S. R. Dahal;Jacob L Lewellen;Shine Ayyappan;B. Chaudhary;Viswanath Nukala;Smita Mohanty