Oriented-sample NMR enables one to study structure and function of membrane proteins under their native-like conditions. However, relatively low sensitivity of this technique still constitutes a major roadblock towards the applications of the method to larger polytopic membrane proteins. Here we show that a combination of membrane-embedded radicals and the repetitive cross-polarization scheme allows one to boost sensitivity by up to a factor three for Pf1 coat protein reconstituted in magnetically aligned bicelles. Such a sensitivity enhancement makes it possible to acquire 15N-15N correlation spectra at a fraction of the previous experimental times and establish spectroscopic assignment in both flipped and unflipped bicelles. Moreover, a new class of experiments based on selective excitation and spectral editing is now possible that allows one to initiate spectroscopic assignment in crowded spectral regions. Results of our recent efforts on constructing a room-temperature dynamic nuclear polarization (DNP) apparatus for macroscopically aligned samples will be presented.

Sensitivity enhancement in solid-state NMR of oriented membrane proteins

254th National Meeting (2017)

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