Acquisition of a 500 MHz NMR Spectrometer for Structural Analysis of Biological Macromolecules

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We propose to establish a Biomolecular NMR Center in Cornell's Biotechnology Building through acquisition of a state-of-the-art 600 MHz high resolution multinuclear NMR spectrometer, to be dedicated for research and training in structural biology. The interdisciplinary biophysics program at Cornell is highly committed to studies of macromolecular structure. This group has established internationally recognized facilities and faculty for X-ray crystallography, theoretical/computational studies of acromolecular structure and dynamics, and biochemical and molecular genetic studies of protein function. The proposed 600 MHz NMR spectrometer will complement existing resources by providing capabilities for solution studies of structure and dynamics. A single 500 MHz NMR spectrometer which is not equipped with the latest technological advances (e.g. fourth radio frequency channel, XYZ pulsed field gradient amplifiers and probes, 10 mm triple resonance probe and enhanced room temperature shims) is housed in Cornell's Chemistry NMR Facility. This machine is routinely booked to full capacity with an extensive user base composed of both "small molecule" and "macromolecule" users. Past progress in solution NMR studies of proteins at Cornell has occurred largely through utilization of outside resources either in the laboratories of collaborators or at public facilities. Over the past decade, triple- and quadruple-resonance multidimensional NMR spectroscopy has become a critical component of modern structural biology. In order to keep pace in this accelerating field of research, and to prepare our undergraduate and graduate students for careers in one of the fastest growing areas of science, this technology must be brought to Cornell. The Director of the Biomolecular NMR Center will be Professor Linda Nicholson who joined the Cornell faculty last fall. Professor Nicholson brings up-to-date expertise in the application of multi-dimensional NMR to dynamical and stru ctural studies of proteins, as evidenced by her recent work on the HIV-l protease performed at the NIH. She brings knowledge of the latest double-, triple- and quadruple- resonance techniques for application to biological macromolecules. This expertise is highly complemented at Cornell by the long-established programs of Harold Scheraga and Robert Oswald that extensively utilize multi-dimensional NMR for structural studies of proteins. progress in these research groups has been made only Day to day direction of the facility will be provided by a staff scientists who will assist and train users, provide technical support and facilitate collaborative interactions. External review of the NMR facility will be provided by the visiting committee described in section 5d. Allocation of resources will be decided by a committee that consists of the director, the staff scientist, one major user and one other user. Scheduling of spectrometer time will be the responsibility of the staff scientist. Cost sharing for this project will be provided by funds from the College of Arts and Sciences and from the Program in Molecular Structure. Nicholson, Oswald and Scheraga comprise the "critical mass" of major users that will launch the first structural and dynamics studies in the proposed Biomolecular NMR Center. Several additional faculty members, including Begley, Jelinski, Rodriguez, and Feigenson, have a demonstrated expertise in solution NMR spectroscopy, and will comprise a strong core of additional users. Research activities encompass investigation of two ubiquitous biophysical switching mechanisms (protein phosphorylation and GTP binding, Nicholson and Oswald), protein folding and protein- protein interactions (Scheraga), the basic forces that hold proteins together (observing the effects of pressure on protein flexibility, Ealick), basic forces governing lipid behavior (Feigenson), the role of structure and dynamics in mechanisms of enzyme catalysis (Begley), and the bi ophysical basis of regulation of gene transcription (Lis and Tye), the biophysical basis of high-performance biopolymers (Jelinski), and interactions between bioactive natural products and DNA (Rodriguez). Methodologies include gradient- enhanced multidimensional experiments to obtain sequential assignments, distance restraints and dihedral angle restraints for structure determination and relaxation parameters for protein dynamics. Studies of larger and less soluble systems will also be possible with the requested instrumentation. In addition to scientific research, the NMR facility will provided educational opportunities through formal courses and training sessions, through graduate and post-graduate research projects and through independent studies and field trips for undergraduate students.