Measurement of 14N quadrupole couplings in biomolecular solids using indirect-detection 14N solid-state NMR with DNP.

The quadrupolar interaction experienced by the spin-1 14N nucleus is known to be extremely sensitive to local structure and dynamics. Furthermore, the 14N isotope is 99.6% naturally abundant, making it an attractive target for characterisation of nitrogen-rich biological molecules by solid-state NMR. In this study, dynamic nuclear polarization (DNP) is used in conjunction with indirect 14N detected solid-state NMR experiments to simultaneously characterise the quadrupolar interaction at multiple 14N sites in the backbone of the microcrystalline protein, GB3. Considerable variation in the quadrupolar interaction (>700 kHz) is observed throughout the protein backbone. The distribution in quadrupolar interactions observed reports on the variation in local backbone conformation and subtle differences in hydrogen-bonding; demonstrating a new route to the structural and dynamic analysis of biomolecules.

Uniform illumination of optically dense NMR samples.

We demonstrate a simple, inexpensive method for in situ laser illumination of NMR samples using a stepwise tapered optical fibre to deliver light uniformly along the axis of a 5 mm NMR tube. The optical path length of the incident light inside the sample is about 3 mm, allowing efficient illumination of optically dense samples. The degradation in spectral resolution and the reduction in filling factor are both minimal. Probe modifications are not required.

Chemically amplified 19F-1H nuclear Overhauser effects.

Chemically induced dynamic nuclear polarisation (CIDNP) is explored as a source of nuclear hyperpolarisation in heteronuclear Overhauser effect experiments. A photochemical reaction proceeding through a radical pair intermediate is used to enhance (19)F nuclear magnetisation in 3-fluorotyrosine by more than an order of magnitude with a corresponding increase in the amplitudes of (19)F-(1)H cross-relaxation and cross-correlation effects. The reactions employed are cyclic and leave the sample chemically unchanged. The potential for enhancing the sensitivity of heteronuclear NOEs in (19)F-labelled proteins is discussed.