CAREER: Structure and Dynamical Mechanisms of Membrane Transport Proteins: New Vistas Via Ultra-stable Force Microscopy

为了使细胞正常运作，一些蛋白质必须穿过细胞膜，而另一些蛋白质必须折叠并插入细胞膜。这些复杂的过程是如何在分子尺度上实现的？涉及哪些力量？蛋白质需要多长时间来重新排列自己？该研究项目旨在通过直接观察单个蛋白质在通过膜时的排列和运动来阐明这些问题。传统上用于细胞研究的技术具有固有的挑战。因此，关于与这些关键生物过程相关的复杂分子运动的详细知识很少。PI已经开发并将采用一种新工具-超高精度显微镜-直接研究这些相互作用。这种基于力的独特仪器有可能实现蛋白质和膜在自然状态下（室温下在水中）的原子级精确测量。这些测量，结合计算机建模和传统的生化实验，将阐明蛋白质分泌和膜蛋白折叠的基本方面。更广泛的影响分子生物学，纳米技术和精密测量的接口提供了一个丰富的跨学科的媒介，将广泛提高这个项目的影响。该项目的教育和外联活动旨在满足从小学到研究生院再到成人学习的整个教育领域的迫切需要。这些活动包括：为旨在改善密苏里州小学教育的教师培训项目编写科学教育课程;继续指导高中和本科生;建立激励和培训机制，以增加研究生外联活动的影响;通过互动式公开演讲承诺与普通观众直接对话。总的来说，这些教育和外联活动旨在接触所有年龄和背景的广泛个人。他们的总体目标是促进科学素养，并传达执行尖端科学研究的兴奋。

Intellectual Merit In order for a cell to operate properly, some proteins must pass through membranes while other proteins must fold and insert themselves into membranes. How are these complex processes achieved at the molecular scale? What are the forces involved? How long does it take for the proteins to rearrange themselves? This research project aims to shed light on these issues by directly observing the arrangements and motions of individual proteins as they negotiate membranes. Techniques that are traditionally used in cellular studies have inherent challenges. As a result, there is little detailed knowledge regarding the complex molecular motions associated with these critical biological processes. The PI has developed and will employ a new tool- an ultra-high precision microscope- to directly study these interactions. This unique instrument, which is based on force, offers the potential to achieve atomically precise measurements of proteins and membranes in their natural state (in water at room temperature). These measurements, in conjunction with computer modeling and traditional biochemical experiments, will elucidate fundamental aspects of protein secretion and membrane protein folding. Broader Impacts The interface of molecular biology, nanotechnology and precision measurement provides a rich interdisciplinary medium that will broadly enhance the impact of this project. The educational and outreach activities of this project aim to address pressing needs across the educational spectrum, from elementary through graduate school to adult learning. These activities include: writing science education curriculum for a teacher training program aimed at improving elementary education in the state of Missouri; continued mentoring of high school and undergraduate students; creating incentives and training mechanisms to increase the impact of graduate student outreach activities; and entering direct dialogue with general audiences through interactive public speaking commitments. Collectively, these educational and outreach activities are designed to reach a broad range of individuals of all ages and backgrounds. Their overall objective is to promote scientific literacy and to convey the excitement of performing cutting edge scientific research.