Abstract
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 8 mL of fully isotope-enriched cell culture. Our approach offers effective means to study proteins with atomic resolution in increasingly native concentrations and environments.
| Original language | English |
|---|---|
| Journal | Angewandte Chemie - International Edition |
| Volume | 55 |
| Issue | 36 |
| Pages (from-to) | 10746-10750 |
| Number of pages | 5 |
| ISSN | 1433-7851 |
| DOIs | |
| Publication status | Published - 26 Aug 2016 |
| Externally published | Yes |
Keywords
- cell lysates
- NMR spectroscopy
- proteins
- structural biology
- structure elucidation