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

通过固态 NMR 均匀标记蛋白质的归属和结构表征

• 批准号: 0749381
• 负责人: Ann McDermott
• 金额: $ 72万
• 依托单位: Columbia University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-15 至 2013-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0749381&HistoricalAwards=false
• 关键词: Assignment; Structural; Characterization; Uniformly; Labeled

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.

本项目的目标是开发用于蛋白质结构测定的新的固体核磁共振方法。在之前的授权期内，首席研究人员已经完成了微晶人泛素的特定部位的核磁共振指定，包括流动区域。此外，她还发起了对噬菌体PF1的螺旋外壳蛋白的指定。她的团队还仅根据固体核磁共振数据确定了泛素的3D结构，达到中等分辨率。在这个项目中，首席研究人员将进一步完善这一结构，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希望的结构约束：基于顺磁标签效应的长程约束和基于对单轴排列样品的研究的角度约束。这些实验有可能在各种生物聚合物研究中带来新的见解，并导致确定许多具有基础和医学意义的不溶性和非晶态蛋白质体系的结构。这个项目更广泛的影响是对学生和博士后研究人员的教育，以及他们在劳动力中的最终贡献。高中生和本科生每年都会通过一系列正在进行的大学项目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：中学女生科学计划、暑期高中科学计划、针对研究生计划中代表人数较少的少数族裔候选人的外展计划、夏季本科生研究奖学金、针对科学教师的暑期研究计划“科学与数学伙伴关系”、哥伦比亚大学的NSF REU计划以及支持哥伦比亚大学本科理科专业的Rabi学者计划。首席研究员的实验室已经与所有这些项目以及一个中学女生的科学项目建立了合作伙伴关系，在这个项目中，她的研究生在计划和运行一年一度的“女孩科学日”(http://www.columbia.edu/cu/news/04/12/girlScienceDay.html).活动中发挥着领导作用。关于哥伦比亚大学新的科学学院(与纽约市学校联合运营)，计划为教师和学生提供一套更广泛的外展项目，首席调查员将参与这些项目的设计，因为她是负责学术规划和科学倡议的副校长的行政职责。