RUI:Test and Measurement Strategies for the Development of Biological Nanovalves

在化学、生物工程、环境和运输系统系化学测量和成像计划的支持和化学、生物工程、环境和运输系统部门界面工程计划的共同资助下，Daniel Burden博士、Lisa Keranen-Burden博士和他们在惠顿学院化学系的团队正在开发一套新的基于激光的工具，将超灵敏显微镜与电子测量和蛋白质工程相结合，以监控带电物种在纳米孔中的移动，其中蛋白质毒素用作纳米阀门。他们的研究寻求以下问题的答案：允许什么分子大小、形状和电荷状态流经纳米级阀门内部，以及必须对纳米阀门进行哪些化学修饰才能使其根据需要从完全开启状态切换到完全关闭状态。所获得的洞察力可能会对广泛的应用有用，包括选择性地进入细胞内部以对抗感染、纳米级分离和化学传感器。该研究团队还在努力通过投资于本科生研究来扩大高级STEM学位合格候选人的规模和多样性。研究的要素被整合到本科课程、暑期研究机会和周围社区的公共推广活动中。离子通道电生理学中的测量方法传统上被用来监测带电物种在纳米孔中的运动。然而，许多翻译事件是电气静默的，无法检测到。此外，了解可以传输(或阻止)的物种的大小、形状、电荷状态和流速，以及它们与纳米阀浇口机制的关系，对于开发该领域未来的应用至关重要。负担小组正在研究是否可以对阿尔法-HL毒素蛋白进行化学修饰，以可逆地停止或允许按需进行分子流动。他们还假设，当触发阀门关闭时，运输分子的大小和速率将发生显着变化。他们正在开发基于广域荧光显微镜、单分子共聚焦显微镜和离子通道电生理学的测量工具，以测试一系列纳米阀结构，最终目的是实时监测基于芯片的双层设备中分子在脂膜上的传输。PI正在与多名本科生合作，特别是接触来自芝加哥地区的第一代进入大学的非洲裔美国人或拉美裔学生。PIS还积极与外展出版商合作，改善向公众传达研究成果。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry and co-funding from the Interfacial Engineering Program in the Division of Chemical, Bioengineering, Environmental, and Transport Systems, Dr. Daniel Burden, Dr. Lisa Keranen-Burden, and their group in the Chemistry Department at Wheaton College are developing a new set of laser-based tools that combine ultrasensitive microscopy with electrical measurements and protein engineering to monitor the movement of charged species through nanopores wherein protein toxins are used as nanovalves. Their studies seek answers to questions such as what molecular sizes, shapes, and charge states are allowed to flow through the nanoscale valve interior, and what chemical modifications to the nanovalve are necessary to make it switchable on demand from the fully-on state to the fully-off state. The insights gained will likely be useful for a broad range of applications, including selective access to the interior of cells to fight infections, nanoscale separations, and chemical sensors. The research team is also working to enhance the size and diversity of the pool of qualified candidates for advanced STEM degrees by investing in undergraduate research. Elements of the research are integrated into the undergraduate curriculum, summer research opportunities, and public outreach activities in the surrounding community. Measurement methods in ion-channel electrophysiology have traditionally been used to monitor the movement of charged species through nanopores. However, many translation events are electrically silent and cannot be detected. Furthermore, an understanding of the sizes, shapes, charge states, and flow rates of species that can be transported (or blocked), as well as their relationship to nanovalve gating mechanisms, is critically important for developing future applications within the field. The Burden group is investigating whether the alpha-HL toxin protein can be chemically modified to reversibly halt or permit molecular flow on demand. They also hypothesize that the size and rate of transported molecules will shift markedly upon triggered valve closure. They are developing measurement tools based on wide-field fluorescence microscopy, single-molecule confocal microscopy, and ion-channel electrophysiology to test a range of nanovalve constructs, with the ultimate aim of monitoring molecular transport across lipid membranes in a chip-based bilayer apparatus in real time. The PIs are working with multiple undergraduate students, reaching out particularly to first-generation college-bound African-American or Latino students from the Chicago area. The PIs are also actively working with outreach publishers to improve communicating research outcomes to the public.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.