CAREER: Biosensor Development for Probing Nanoscale Topology in Neurotransmission

1452057 -Knowles测量生物分子的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，仿生系统提供了简单的平台 为 结合 膜 蛋白 成 生物传感 应用. 在 在这些应用中，膜蛋白能够感知它们的局部环境，例如化学成分和膜形状，而这些因素影响蛋白质的功能。到目前为止，很少有人关注这些传感器的纳米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识别细胞和生物分子如何受到纳米结构材料影响的生物传感器。 具体而言，蛋白质参与神经元信号的传输和激素的分泌将被表征。研究与教育的整合将通过在暑期工程夏令营中对8 - 12年级学生的推广，在暑期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接口设计新颖的课程来实现。与生物系统接口的材料工程越来越需要了解纳米级拓扑结构的细胞和分子反应。这个CAREER奖的重点是设计两个生物传感器，用于探测纳米结构材料和蛋白质功能之间的关系。第一个传感器将模拟细胞内质膜，具有膜曲率和化学组成的可调区域。第二个将提供一个模板，将膜曲率引入活细胞，在那里可以评估对曲率的分子反应。这两种传感器将用于表征膜形状和神经传递之间的关系，这是一个引起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，以促进栓系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再生领域做出贡献。通过使用超分辨率荧光显微镜技术，单粒子跟踪，并结合共聚焦荧光原子力显微镜，管理纳米结构膜上的蛋白质分选的原则将被确定。 这些原则将在未来的生物传感器和与细胞接口的材料的设计中是有用的。这项研究与教育的整合将通过推广到8 - 12年级的学生在暑期工程营，在夏季研究计划期间培训当地教育工作者，并在生物化学和工程的界面设计新颖的课程。

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.