Acquisition of a 600 MHz NMR Cryoprobe

University Of Md Biotechnology Institute, Baltimore MD

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Abstract

The Center for Advanced Research in Biotechnology (CARB), a joint research Institute of the University of Maryland Biotechnology Institute (UMBI) and the National Institute of Standards and Technology (NIST), is applying for funds to assist in the purchase of a Bruker MHz Nuclear Magnetic Resonance (NMR) TXI 5 mm sample diameter cryoprobe system, or the equivalent. The proposed 600 MHz NMR cryoprobe system will be used primarily by NIH funded researchers at CARB to investigate the structure and dynamics of biological macromolecules using multi-dimensional homo- and heteronuclear NMR methods. Currently, the research programs of five principle investigators at CARB, who comprise the primary NMR user group, would benefit significantly by the acquisition of the proposed 600 MHz NMR cryoprobe system include: 1) Membrane-targeting Calcium Sensors in Vision (Ames); 2) Protein Folding and Stability of Subtilisin (Bryan); 3) Structural Basis of RNA Stem-Loop Kissing Complexes (Marino); 4) Frm Genomic Sequences in Protein Structure and Function: Structural Data from NMR Spectroscopy (Orban); 5) Folding and Assembly in Visual Rhodopsin (Ridge/Brabazon). The proposed 600 MHz cryoprobe system will allow NMR data to be acquired with a sensitivity that is approximately a factor of 4 greater than conventional 600 MHz NMR systems. We anticipate that it will therefore open up the possibility to acquire NMR data for samples/experiments which are currently impractical to measure due to either inherent weakness of the NMR correlation, low solubility and/or limited sample availability. Additionally, it will dramatically reduce measurement time by approximately a factor of 16 for conventional samples/experiments., thereby accelerating the NIH funded research programs. There are important considerations to our proposal. First, access to a 600 MHz NMR cryoprobe system is currently not available either within the UMBI system or NIST. The proposed instrumentation will therefore not only be a valuable resource for CARB researchers, but also for scientists at the four other research centers of UMBI [i.e. The Institute for Human Virology (IHV), Center for Agricultural Biotechnology (CAB), Center for Marine Biotechnology (COMB) and the Medical Biotechnology enter MDC)] and a NIST who may need to use its unique capabilities in the studies of biological macromolecules. Second, the proposed 600 MHz cryoprobe system will be housed in our recently constructed shared NMR laboratory and maintained with committed infrastructure funds from CARB. Third, a new facility manager (Sari) has also been hired to maintain the 600 MHz NMR cryoprobe system as part of the CARB NMR facility.