NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: Microcrystals, sedimented assemblies, and amyloid fibrils.
- Stanek, J.
- Andreas, L.
- Jaudzems, K.
- Cala, D.
- Lalli, D.
- Bertarello, A.
- Schubeis, T.
- Akopjana, I.
- Kotelovica, S.
- Tars, K.
- Pica, A.
- Leone, S.
- Picone, D.
- Xu, Z.
- Dixon, N.
- Martinez, D.
- Berbon, M.
- El Mammeri, N.
- Noubhani, A.
- Saupe, S.
- Habenstein, B.
- Loquet, A.
- Pintacuda, G.
DOIAbstract
We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100–111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα–Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration
