CAREER: Biosensor Development for Probing Nanoscale Topology in Neurotransmission

职业：用于探测神经传递中纳米级拓扑的生物传感器开发

• 批准号: 1452057
• 负责人: Michelle Knowles
• 金额: $ 50万
• 依托单位: University of Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-15 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1452057&HistoricalAwards=false
• 关键词: CAREER; Biosensor; Development; Probing; Nanoscale; CAREER; Biosensor; Development; Probing; Nanoscale

1452057 - KnowlesSensors that measure biological molecules are essential for medical diagnostics and drug discovery. These sensors often rely on the measurement of membrane proteins, which are targets for drug discovery. On the other hand, biomimetic systems provide simple platforms for incorporating membrane proteins into biosensing applications. In these applications, membrane proteins are able to sense their local environment, such as the chemical composition and membrane shape, and these factors affect protein function. Until now little attention has been given to the nanoscale structure of these sensors and how nanometer-sized features affect proteins. The main focus of this research is to design biosensors that can be used to identify how cells and biomolecules are affected by nanostructured materials. Specifically, proteins involved with the transmission of neuronal signals and the secretion of hormones will be characterized. The integration of research with education will take place through outreach to 8th-12th grade students in a summer engineering camp, training of local educators during a summer research program, and the design of novel courses at the interface of biochemistry and engineering.The engineering of materials that interface with biological systems increasingly requires an understanding of cellular and molecular responses to nanoscale topology. The focus of this CAREER Award is to design two biosensors that will be used to probe the relationship between nanostructured materials and protein function. The first sensor will mimic the intracellular plasma membrane with tunable regions of membrane curvature and chemical composition. The second will provide a template to introduce membrane curvature into live cells, where the molecular response to curvature can be assessed. Both sensors will be used to characterize the relationship between membrane shape and neurotransmission, a biological process that gives rise to extreme changes in membrane topology. Neurotransmission relies on the proper recruitment and function of SNARE proteins to facilitate the fusion of the tethered vesicle membrane with the plasma membrane. SNARE-mediated membrane fusion is essential for membrane repair, growth cone formation, axon extension, and synapse formation. Our work aspires to contribute to the field of neuroregeneration by identifying nanoscale features that drive membrane fusion. By using super-resolution fluorescence microscopy techniques, single particle tracking, and combined confocal fluorescence-atomic force microscopy, principles that govern protein sorting on nanostructured membranes will be identified. These principles will be useful in the future design of biosensors and materials that interface with cells. The integration of this research with education will take place through outreach to 8th-12th grade students in a summer engineering camp, training of local educators during a summer research program, and the design of novel courses at the interface of biochemistry and engineering.

1452057-测量生物分子的知识传感器对于医学诊断和药物发现至关重要。这些传感器通常依赖于膜蛋白的测量，这些膜蛋白是药物发现的靶标。另一方面，仿生系统提供了简单的平台，用于将膜蛋白掺入生物传感应用中。 在这些应用中，膜蛋白能够感知其局部环​​境，例如化学成分和膜形状，这些因素会影响蛋白质功能。到目前为止，几乎没有关注这些传感器的纳米级结构，以及纳米尺寸的特征如何影响蛋白质。这项研究的主要重点是设计可用于确定细胞和生物分子如何受纳米结构材料影响的生物传感器。 具体而言，将表征与神经元信号传递和激素分泌有关的蛋白质。研究的整合将通过向夏季工程营地的8年至12年级的学生推广，在夏季研究计划中对当地教育工作者的培训以及在生物化学和工程学的界面上设计新课程的培训。该职业奖的重点是设计两个生物传感器，这些生物传感器将用于探测纳米结构材料和蛋白质功能之间的关系。第一个传感器将与膜曲率和化学成分的可调区域模拟细胞内质膜。第二个将提供一个模板将膜曲率引入活细胞中，在该细胞中可以评估分子对曲率的响应。这两个传感器都将用于表征膜形和神经传递之间的关系，这是一种生物学过程，从而导致膜拓扑的极端变化。神经传递依赖于SNARE蛋白的适当募集和功能来促进束缚囊泡膜与质膜的融合。 SNARE介导的膜融合对于膜修复，生长锥形成，轴突延伸和突触形成至关重要。我们的工作渴望通过识别驱动膜融合的纳米级特征来为神经发生的领域做出贡献。通过使用超分辨率荧光显微镜技术，单个颗粒跟踪以及共聚焦荧光原子力显微镜，将确定控制纳米结构膜上蛋白质分类的原理。 这些原理在将来的生物传感器和与细胞接口的材料的未来设计中很有用。这项研究与教育的融合将通过向夏季工程营地的8至12年级学生推广，在夏季研究计划中对当地教育工作者的培训以及在生物化学和工程学界面上设计新颖课程的培训。