Premium
Selective Protein Hyperpolarization in Cell Lysates Using Targeted Dynamic Nuclear Polarization
Author(s) -
Viennet Thibault,
Viegas Aldino,
Kuepper Arne,
Arens Sabine,
Gelev Vladimir,
Petrov Ognyan,
Grossmann Tom N.,
Heise Henrike,
Etzkorn Manuel
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201603205
Subject(s) - hyperpolarization (physics) , chemistry , nuclear magnetic resonance spectroscopy , polarization (electrochemistry) , lysis , nuclear magnetic resonance , biophysics , biochemistry , stereochemistry , biology , physics
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.