-- EB002026 This proposal is focused on the development of high frequency dynamic nuclear polarization (DNP) nuclear magnetic resonance (NMR) to enhance sensitivity in NMR based structural biology experiments. 1 CW DNP at 527 GHz/800 MHz: We plan to complete development of a 527 GHz/800 MHz DNP spectrometer. It will involve (1) completion of low temperature (LT), 3 channel MAS probes, (2) the cryogenics for MAS at 80 K, (3) corrugated waveguide, and (4) interfacing these components with a 527 GHz tunable gyrotron. This will be the sole 527 GHz/800 MHz DNP instrument available in North America. 2. Applications to Peptides and Proteins: We plan to continue application of DNP to studies of proteins The specific aim is to optimize the polarizing agents and NMR methodology for 2H,13C,15N labeled amyloids, in particular tau, and to determine the structure of tau fibrils and PET imaging binding sites. We will also determine the structure of H217O in fibrils with 17O DNP experiments on TTR and GNNQQNY 2 High B0 Cross Effect (CE) Polarizing Agents: We plan to develop new polarizing agents for CE DNP at 600-800 MHz. They will use larger e--e- dipole couplings, trityl-TEMPO biradicals, and narrow line radicals with different isotropic g-values that satisfy the cross effect matching condition. 3 Overhauser Effect (OE) DNP in Solids: We plan to develop narrow line trityl radicals with large 1H hyperfine couplings that mediate OE DNP. The temperature, ?1S and ?0S dependence of these radicals will be studied. In addition we will study specifically 2H labeled BDPA to determine the mechanism of the OE. 4 Pulsed DNP Experiments:
The specific aim i s to move pulsed DNP to 250 GHz using an EPR spectrometer that is centered around a fast AWG, a 600-1000 watt 250 GHz gyroamplifier and a MAS probe with a TE011 resonator and 1.3 mm rotor. We will investigate NOVEL, off resonance NOVEL, the frequency sweep integrated solid effect (FS-ISE), and time optimized DNP (TOP DNP) as approaches for time domain polarization transfer. We plan to develop adiabatic versions of the latter. 6 1H Detected DNP: We propose to develop methods for 1H detected DNP of 13C, 15N and 17O resonances using a helium recirculation system that will permit ?r/2?>100 kHz at 90 K. The system will initially use 1.3 and 0.7 rotors from ZrO2 and we will develop laser machined diamond rotors that will spin at >100 kHz. The recirculation system will also be useful for spinning at ambient temperatures at >200 kHz.

Public Health Relevance

The research proposes to use dynamic nuclear polarization to improve the sensitivity in magic angle spinning NMR experiments, and the higher sensitivity will lead to molecular structures with higher precision and could have a profound impact on structural biology, medical imaging and solution NMR. In addition, we propose to develop new polarizing agents for DNP, new equipment for the experiments, and new time domain methods for more rapid data acquisition. The experiments will be applied to a variety of model and unknown systems whose structure we will determine.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
9R01GM132997-31
Application #
9738551
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Krepkiy, Dmitriy
Project Start
1987-04-01
Project End
2023-05-31
Budget Start
2019-06-01
Budget End
2020-05-31
Support Year
31
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Type
Organized Research Units
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02142