Funding is requested for the purchase of three essential items that will be used with a 800/89 magnet: (1) a spectrometer console, (2) room temperature shims and (3) a magic angle spinning (MAS) probe with a 1.3 mm stator rotor system. The magnet was delivered in February and is presently at field. In the immediate future the spectrometer will be operated as a high field MAS NMR with low temperature capabilities. However, in the long term it will be converted to an 800 MHz/527 GHz DNP instrument. With funds from another grant we purchased a probe capable of performing experiments in the 90-300 K range and will be used to examine a variety of temperature dependent phenomena in cryoprotected membrane protein and amyloid protein samples. The probe has a vacuum jacketed dewar, sample eject capabilities, and includes a waveguide for dynamic nuclear polarization (DNP) experiments at 527 GHz. Research projects include the following investigations of membrane and amyloid proteins: (1) bacteriorhodopsin (bR) photocycle intermediates in native membranes and (2) nanodiscs, (3) the voltage dependent anion channel (VDAC) in lipid bilayers, (4) the M2 protein from influenza virus, (5) gas vesicles which form a proteinaceous membrane, and amyloid fibrils formed from (6) phosphatidyl-inositol-3-kinase SH3 domain (PI3- SH3), (7) beta-2-microglobulin associated with dialysis related amyloidosis, and (8) the CsgA protein, from Curli. In addition, this will be a prototype instrument performing DNP experiments at 800/527 and will (9) be used to test new biradical polarizing agents, (10) develop high field 17O MAS experiments and (11) new techniques for dipole recoupling at high ?r/2?. The 800/89 will be part of a cluster of instrumentation that serves a community of scientist performing high field biological NMR in the Boston area and North America. It will alleviate a pressing need for additional capabilities for low temperature and high field DNP/ MAS experiments. Further, the instrument will have a significant impact on research since it will employ a number of innovative technologies that are not available elsewhere. For example, the combination of the LT-MAS probe with the 800/89 will be one of the few widebore, low temperature solid state NMR instruments available worldwide and the addition of a 527 GHz gyrotron will make the system a unique DNP spectrometer in North America. It will therefore fill an important void in structural biology.
The 527 GHz gyrotron and 70 kHz MAS probe operating on the 800/89 magnet will be used to investigate the structure of membrane and amyloid proteins. Some of the systems are of biophysical interest and others are associated with amyloidiogenic disease. Thus, the instrumentation will be used in structural investigations of proteins associated with basic protein biophysics and others for particular human diseases
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