title: Selective High-Resolution Detection of Membrane Protein-Ligand Interaction in Native Membranes Using Trityl-Nitroxide PELDOR.
abstract: The orchestrated interaction of transmembrane proteins with other molecules mediates several crucial biological processes. Detergent solubilization may significantly alter or even abolish such hetero-oligomeric interactions, which makes observing them at high resolution in their native environment technically challenging. Dipolar electron paramagnetic resonance (EPR) techniques such as pulsed electro-electron double resonance (PELDOR) can provide very precise distances within biomolecules. To concurrently determine the inter-subunit interaction and the intra-subunit conformational changes in hetero-oligomeric complexes, a combination of different spin labels is required. Orthogonal spin labeling using a triarylmethyl (TAM) label in combination with a nitroxide label is used to detect protein-ligand interactions in native lipid bilayers. This approach provides a higher sensitivity and total selectivity and will greatly facilitate the investigation of multimeric transmembrane complexes employing different spin labels in the native lipid environment.
authors: Institut für physikalische und theoretische Chemie und Biomolekulares Magnetresonanz Zentrum, Universität Frankfurt, Max-von-Laue-Strasse 7, 60438, Frankfurt am Main, Germany. joseph@epr.uni-frankfurt.de.
authors: Benesh
authors: 0000-0003-4968-889X
authors: B
authors: Joseph
authors: N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry SB RAS, Novosibirsk, 630090, Russia.|Novosibirsk State University, Novosibirsk, 630090, Russia.
authors: Victor M
authors: 0000-0003-4975-9993
authors: VM
authors: Tormyshev
authors: N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry SB RAS, Novosibirsk, 630090, Russia.
authors: Olga Yu
authors: OY
authors: Rogozhnikova
authors: Institut für physikalische und theoretische Chemie und Biomolekulares Magnetresonanz Zentrum, Universität Frankfurt, Max-von-Laue-Strasse 7, 60438, Frankfurt am Main, Germany.
authors: Dmitry
authors: D
authors: Akhmetzyanov
authors: N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry SB RAS, Novosibirsk, 630090, Russia.
authors: Elena G
authors: EG
authors: Bagryanskaya
authors: Institut für physikalische und theoretische Chemie und Biomolekulares Magnetresonanz Zentrum, Universität Frankfurt, Max-von-Laue-Strasse 7, 60438, Frankfurt am Main, Germany. prisner@chemie.uni-frankfurt.de.
authors: Thomas F
authors: TF
authors: Prisner
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D008024:Ligands; D008565:Membrane Proteins / Q000737:chemistry*; D008968:Molecular Conformation; D009589:Nitrogen Oxides / Q000737:chemistry; D013113:Spin Labels
chemical_list: D008024:Ligands; D008565:Membrane Proteins; D009589:Nitrogen Oxides; D013113:Spin Labels; C039900:nitroxyl
keywords: EPR; PELDOR or DEER; membrane proteins; spin labeling; trityl