CAREER: Biosensor Development for Probing Nanoscale Topology in Neurotransmission

1452057-测量生物分子的知识传感器对于医学诊断和药物发现是必不可少的。这些传感器通常依赖于膜蛋白的测量，而膜蛋白是药物发现的目标。另一方面，仿生系统提供了将膜蛋白整合到生物传感应用中的简单平台。在这些应用中，膜蛋白能够感知其局部环境，如化学成分和膜形状，这些因素影响蛋白质的功能。到目前为止，对这些传感器的纳米级结构以及纳米级特征如何影响蛋白质的关注很少。这项研究的主要重点是设计生物传感器，可以用来识别细胞和生物分子如何受到纳米结构材料的影响。具体地说，涉及神经元信号传递和激素分泌的蛋白质将被描述。研究与教育的结合将通过在暑期工程夏令营接触8-12年级的学生，在暑期研究计划期间对当地教育工作者进行培训，以及在生物化学和工程的界面上设计新的课程来进行。与生物系统连接的材料工程越来越需要了解细胞和分子对纳米级拓扑的反应。这一职业奖的重点是设计两个生物传感器，用于探索纳米结构材料和蛋白质功能之间的关系。第一个传感器将模拟细胞内的质膜，具有膜曲率和化学成分的可调区域。第二个将提供一个模板，将膜曲率引入活细胞，在那里可以评估分子对曲率的反应。这两个传感器将被用来表征膜形状和神经传递之间的关系，这是一个导致膜拓扑结构极端变化的生物过程。神经传递依赖于SNARE蛋白的适当募集和功能，以促进拴系的囊泡膜与质膜的融合。SNARE介导的膜融合是膜修复、生长锥形成、轴突延伸和突触形成所必需的。我们的工作致力于通过识别驱动膜融合的纳米级特征来为神经再生领域做出贡献。通过使用超分辨率荧光显微镜技术、单颗粒跟踪和共聚焦荧光-原子力显微镜相结合的技术，将确定在纳米结构膜上管理蛋白质分类的原则。这些原理将在未来生物传感器和与细胞连接的材料的设计中有用。这项研究与教育的结合将通过在暑期工程夏令营接触8-12年级的学生，在暑期研究计划期间对当地教育工作者进行培训，以及在生物化学和工程学的界面上设计新颖的课程来进行。

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.