Investigating Membrane Proteins with Magnetic Resonance Spectroscopy

该奖项将资助迈阿密大学化学和生物化学系的生命过程化学项目资助加里·A·洛根教授研究一种名为Pinholin的膜蛋白的结构和动态性质。新的强大的最先进的技术被用于研究Pinholin，它是一种直接参与细胞死亡的重要的膜结合蛋白。病毒感染周期的最后一步是通过裂解导致细胞死亡，即细胞膜的破裂。洛根教授计划开发实验程序，以表征膜中针孔蛋白活性形式和非活性形式的结构和机制。这一追求使研究生、本科生和来自代表性不足群体的学生能够获得尖端电子顺磁共振(EPR)生物物理技术方面的专门培训。发展起来的新的生物物理方法对研究各种不同的膜蛋白系统普遍有用。该研究项目还促进了迈阿密大学名为DUOS的变革性STEM指导计划的发展，该计划加强了研究生和本科生研究人员之间的协同/互动，以便在博士授予系的教职员工的监督下就研究项目进行合作。来自洛根实验室的一个本科生/研究生团队计划参与这一新项目。该研究项目旨在表征针孔蛋白活性形式和非活性形式的结构、拓扑、动力学和寡聚状态。采用强大的CW-EPR和脉冲EPR光谱技术研究了针孔蛋白的不同构型。要测量自旋标记之间的多个距离，以提供嵌入在膜中的针孔蛋白的重要结构信息。这项研究的信息将为针孔蛋白的机制和细胞裂解提供新的见解，并提供普遍适用于膜蛋白的新方法。该奖项反映了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 this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Professor Gary A. Lorigan from the Department of Chemistry and Biochemistry at Miami University to investigate the structural and dynamic properties of a membrane protein known as pinholin. Novel powerful state-of-the-art techniques are used to study pinholin, which is an important membrane-bound protein that is directly involved in cell death. The last step of the virus infection cycle is cell death through lysis, the breakdown of the cell membrane. Professor Lorigan plan to develop the experimental procedures to characterize the structure and mechanism of the active and inactive forms of pinholin in a membrane. The pursuit allows graduate students, undergraduate students, and students from under-represented groups to obtain specialized training in cutting-edge Electron Paramagnetic Resonance (EPR) biophysical techniques. The new biophysical methods developed are useful generally to study a variety of different membrane protein systems. This research project also enables the development of a transformative STEM mentoring program called DUOS at Miami University that strengthens the synergy/interaction between graduate and undergraduate researchers to collaborate on a research project under the supervision of a faculty member in a Ph.D. granting department. One undergraduate/graduate student team from the Lorigan lab is scheduled to participate in this new program.The research project is conducted to characterize the structure, topology, dynamics, and oligomerization state of the active and inactive forms of pinholin. Powerful CW-EPR and pulsed EPR spectroscopic techniques are used to investigate the different formations of pinholin. Multiple distances are to be measured between spin labels to provide important structural information on the pinholin embedded in a membrane. Information from this study should provide new insight into the pinholin mechanism and cell lysis, and provide novel methodology generally applicable to membrane proteins.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.