Calmodulin-based allosteric switches as a modular platform for peptide biosensors

1158784尼尔·S·乔希所有的生物传感平台都建立在两根柱子上？对特定分析物的特定生化识别以及将该识别转换为易于检测的信号。大多数现有的生物传感技术利用被动与其分析物结合的蛋白质，因此需要浪费的洗涤步骤、专门的试剂和昂贵的检测仪器。PI建议开发一种新的工程蛋白质开关，这种开关可以主动感知基于多肽的分析物，并仅在分析物结合时才直接产生输出信号。智能优点：将传感和输出信号产生的动作结合到单个蛋白质中，将产生适合用于便携式生物传感设备的蛋白质开关，这些设备可以在现场使用，也可以供未经培训的个人使用，从而显著简化生物样品中基于多肽的分析物的量化。更广泛的影响：本提案中开发的技术将作为进一步开发能够快速检测生物样品中的疾病生物标记物或来自环境来源的致病毒素的设备的基础。这个项目的资金将支持一个本科生研究团队参加一个独特的国际生物分子设计研究竞赛(Biood.net)，还将支持一个正在进行的推广计划，以促进来自科学代表性较低社区的当地高中生与Wyss生物启发工程研究所的研究人员之间的互动。该项目由分子和细胞科学系工程和分子生物物理学的CBET计划联合资助。

1158784Neel S. JoshiAll biosensing platforms rest on two pillars ? specific biochemical recognition of a particular analyte and transduction of that recognition into a readily detectable signal. Most existing biosensing technologies utilize proteins that passively bind to their analytes, and therefore require wasteful washing steps, specialized reagents, and expensive instruments for detection. The PI proposes to develop a new class of engineered protein switches that can actively sense peptide-based analytes and directly produce an output signal only when the analyte is bound. Intellectual Merit: Combining the actions of sensing and output signal generation into a single protein will result in protein switches that are suitable for use in portable biosensing devices that can be used in the field and by untrained individuals, significantly simplifying the quantification of peptide-based analytes in biological samples. Broader Impacts: The technology developed in this proposal will serve as a foundation for the further development of devices capable of rapid detection of disease biomarkers in biological samples or pathogenic toxins from environmental sources. Funding for this project will support a team of undergraduate researchers to participate in a unique international research competition for biomolecular design (biomod.net) and also support an ongoing outreach program to promote interaction between local high school students from communities underrepresented in the sciences and researchers at the Wyss Institute for Biologically Inspired Engineering.This Project is jointly funded by CBET program at Engineering and Molecular Biophysics in the Division of Molecular and Cellular Sciences.