Assignment and Structural Characterization of Uniformly Labeled Proteins by Solid State NMR

本项目的目标是开发新的固态核磁共振方法用于蛋白质结构测定。在上一个资助期，主要研究者已经完成了微晶人泛素的位点特异性NMR共振分配，包括移动的区域。此外，她开始了噬菌体Pf1螺旋外壳蛋白的任务。她的团队还仅基于固态NMR数据确定了泛素的3D结构，分辨率适中。 在这个项目中，主要研究者将进一步完善这种结构，并确定噬菌体外壳蛋白寡聚体的结构。 首席研究员还将开发两种新的有前途的结构约束：基于顺磁标签效应的长距离约束，以及基于研究单轴对齐样品的角度约束。这些实验有可能导致新的见解，在各种生物聚合物的研究，并导致许多不溶性和非结晶蛋白质系统的基本和医学重要性的结构的测定。该项目的更广泛影响是学生和博士后研究人员的教育，以及他们对劳动力的最终贡献。高中生和本科生每年都在首席研究员的实验室接受结构生物学和磁共振的培训和接触，通过一些正在进行的大学课程：中学女生科学项目，夏季高中科学项目，针对研究生项目代表性不足的少数民族候选人的外展项目，夏季本科研究奖学金，科学教师暑期研究项目“科学与数学伙伴关系”、哥伦比亚大学的NSF REU项目，以及支持哥伦比亚大学本科科学专业的拉比学者项目。 主要研究者的实验室与所有这些项目合作，并与一个中学女生科学项目合作，她的研究生在计划和举办年度活动“女生科学日”（http：//www.columbia.edu/cu/news/04/12/girlScienceDay.html）中发挥领导作用。在与哥伦比亚大学的新科学学院（与纽约市学校联合运营）的联系中，计划为教师和学生提供一套更广泛的外展计划，首席研究员将参与这些计划的设计，因为她是负责学术规划和科学倡议的副总裁。

The objective of this project is to develop new solid state NMR methods for protein structure determination. In the previous grant period, the principal investigator has completed the site-specific NMR resonance assignments for microcrystalline human Ubiquitin, including mobile regions. In addition, she initiated the assignments for the helical coat protein of bacteriophage Pf1. Her group has also determined the 3D structure of Ubiquitin based on solid-state NMR data alone, to moderate resolution. In this project, the principal investigator will further refine this structure and determine the structure of the bacteriophage coat protein oligomer. The principal investigator will also develop two new kinds of promising structural restraints: long-range constraints based on the effects of paramagnetic tags, and angular restraints based on studying a uniaxially aligned sample. These experiments have the potential to lead to new insights in a variety of biopolymer studies, and to lead to the determination of structures of many insoluble and non-crystalline protein systems of fundamental and medical importance. The broader impact of this project is the education of students and post-doctoral researchers, and their ultimate contributions in the workforce. High school and undergraduate students are trained and exposed to structural biology and magnetic resonance in the principal investigator's laboratory every year, via a number of ongoing university programs: Middle School Girl's Science Program, The Summer High School Science Program, outreach programs targeted at underrepresented minority candidates for the graduate programs, Summer Undergraduate Research Fellowships, the summer research program for science teachers "Science and Math Partnership", Columbia University's NSF REU program, and the Rabi Scholars program, which supports Columbia University's undergraduate science majors. The principal investigator's laboratory has partnered with all of these programs, and with a middle school girl's science program, where her graduate students play a leadership role in planning and running the annual event, "Girls' Science Day" (http://www.columbia.edu/cu/news/04/12/girlScienceDay.html). In connection with Columbia University's new Science School (jointly run with the NYC Schools), a more extensive set of outreach programs for teachers and students is planned, and the principal investigator will be involved in the design of these programs due to her administrative responsibilities as Associate Vice President for Academic Planning and Science Initiatives.