title: Selective Protein Hyperpolarization in Cell Lysates Using Targeted Dynamic Nuclear Polarization.
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.
authors.affiliation: Institute of Physical Biology, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.|Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Wilhelm Jonen Strasse, Jülich, Germany.
authors.forename: Thibault
authors.identifier: 0000-0001-5349-0179
authors.initials: T
authors.lastname: Viennet
authors.affiliation: Institute of Physical Biology, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.|Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Wilhelm Jonen Strasse, Jülich, Germany.
authors.forename: Aldino
authors.identifier: 0000-0003-1733-136X
authors.initials: A
authors.lastname: Viegas
authors.affiliation: Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, 44227, Dortmund, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Kuepper
authors.affiliation: Institute of Physical Biology, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.|Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Wilhelm Jonen Strasse, Jülich, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Arens
authors.affiliation: Faculty of Chemistry and Pharmacy, Sofia University, 1 James Bourchier Blvd., 1164, Sofia, Bulgaria.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Gelev
authors.affiliation: Faculty of Chemistry and Pharmacy, Sofia University, 1 James Bourchier Blvd., 1164, Sofia, Bulgaria.
authors.forename: Ognyan
authors.initials: O
authors.lastname: Petrov
authors.affiliation: Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, 44227, Dortmund, Germany.|VU University Amsterdam, Department of Chemistry and Pharmaceutical Sciences, De Boelelaan 1083, 1081, HV, Amsterdam, The Netherlands.
authors.forename: Tom N
authors.identifier: 0000-0003-0179-4116
authors.initials: TN
authors.lastname: Grossmann
authors.affiliation: Institute of Physical Biology, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.|Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Wilhelm Jonen Strasse, Jülich, Germany.
authors.forename: Henrike
authors.initials: H
authors.lastname: Heise
authors.affiliation: Institute of Physical Biology, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany. manuel.etzkorn@hhu.de.|Institute of Complex Systems (ICS-6), Forschungszentrum Jülich, Wilhelm Jonen Strasse, Jülich, Germany. manuel.etzkorn@hhu.de.
authors.forename: Manuel
authors.identifier: 0000-0002-9796-3246
authors.initials: M
authors.lastname: Etzkorn
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D003497:Cyclic N-Oxides / Q000737:chemistry; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular; D011092:Polyethylene Glycols / Q000737:chemistry; D020005:Propanols / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000737:chemistry / Q000378:metabolism; D051020:bcl-X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: C521769:1-(TEMPO-4-oxy)-3-(TEMPO-4-amino)propan-2-ol; D002247:Carbon Isotopes; D003497:Cyclic N-Oxides; D020005:Propanols; D011506:Proteins; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051020:bcl-X Protein; D011092:Polyethylene Glycols; C000615229:Carbon-13
keywords: NMR spectroscopy; cell lysates; proteins; structural biology; structure elucidation