Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 55, Issue 36, Pages 10746-10750Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201603205
Keywords
cell lysates; NMR spectroscopy; proteins; structure elucidation; structural biology
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Funding
- Major Equipment Initiative [HE3243/4-1]
- Emmy Noether grants of the DFG [ET103/2-1, GR3592/2-1]
- Marie Sklodowska-Curie Grant [660258]
- International NRW Research School iGRASPseed
- AstraZeneca
- Bayer CropScience
- Bayer Health-Care
- Boehringer Ingelheim
- Merck KGaA
- Max Planck Society
- Marie Curie Actions (MSCA) [660258] Funding Source: Marie Curie Actions (MSCA)
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Nuclear magnetic resonance (NMR) spectroscopy has the intrinsic capabilities to investigate proteins in native environments. In general, however, NMR relies on non-natural protein purity and concentration to increase the desired signal over the background. We here report on the efficient and specific hyperpolarization of low amounts of a target protein in a large isotope-labeled background by combining dynamic nuclear polarization (DNP) and the selectivity of protein interactions. Using a biradical-labeled ligand, we were able to direct the hyperpolarization to the protein of interest, maintaining comparable signal enhancement with about 400-fold less radicals than conventionally used. We could selectively filter out our target protein directly from crude cell lysate obtained from only 8mL of fully isotope-enriched cell culture. Our approach offers effective means to study proteins with atomic resolution in increasingly native concentrations and environments.
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