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

Columbia University, New York NY

Investigators

Abstract

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.