Structural response of G protein binding to the cyclodepsipeptide inhibitor FR900359 probed by NMR spectroscopy.

Autor: Bonifer C; Institute of Biophysical Chemistry, Centre of Biomolecular Magnetic Resonance, Goethe University Frankfurt Max-von-Laue Str. 9 60438 Frankfurt Germany glaubitz@em.uni-frankfurt.de., Hanke W; Institute for Pharmaceutical Biology, University of Bonn Nussallee 6 53115 Bonn Germany., Mühle J; Division of Biology and Chemistry, Laboratory of Biomolecular Research, Paul Scherrer Institute Forschungsstr. 111, 5232 Villigen PSI Switzerland., Löhr F; Institute of Biophysical Chemistry, Centre of Biomolecular Magnetic Resonance, Goethe University Frankfurt Max-von-Laue Str. 9 60438 Frankfurt Germany glaubitz@em.uni-frankfurt.de., Becker-Baldus J; Institute of Biophysical Chemistry, Centre of Biomolecular Magnetic Resonance, Goethe University Frankfurt Max-von-Laue Str. 9 60438 Frankfurt Germany glaubitz@em.uni-frankfurt.de., Nagel J; Department of Pharmaceutical & Medicinal Chemistry, Pharmaceutical Institute, University of Bonn An der Immenburg 4 53121 Bonn Germany., Schertler GFX; Division of Biology and Chemistry, Laboratory of Biomolecular Research, Paul Scherrer Institute Forschungsstr. 111, 5232 Villigen PSI Switzerland., Müller CE; Department of Pharmaceutical & Medicinal Chemistry, Pharmaceutical Institute, University of Bonn An der Immenburg 4 53121 Bonn Germany., König GM; Institute for Pharmaceutical Biology, University of Bonn Nussallee 6 53115 Bonn Germany., Hilger D; Department of Pharmaceutical Chemistry, University of Marburg 35037 Marburg Germany daniel.hilger@pharmazie.uni-marburg.de., Glaubitz C; Institute of Biophysical Chemistry, Centre of Biomolecular Magnetic Resonance, Goethe University Frankfurt Max-von-Laue Str. 9 60438 Frankfurt Germany glaubitz@em.uni-frankfurt.de.
Jazyk: angličtina
Zdroj: Chemical science [Chem Sci] 2024 Jul 04; Vol. 15 (32), pp. 12939-12956. Date of Electronic Publication: 2024 Jul 04 (Print Publication: 2024).
DOI: 10.1039/d4sc01950d
Abstrakt: The cyclodepsipeptide FR900359 (FR) and its analogs are able to selectively inhibit the class of G q proteins by blocking GDP/GTP exchange. The inhibitor binding site of G q has been characterized by X-ray crystallography, and various binding and functional studies have determined binding kinetics and mode of inhibition. Here we investigate isotope-labeled FR bound to the membrane-anchored G protein heterotrimer by solid-state nuclear magnetic resonance (ssNMR) and in solution by liquid-state NMR. The resulting data allowed us to identify regions of the inhibitor which show especially pronounced effects upon binding and revealed a generally rigid binding mode in the cis conformation under native-like conditions. The inclusion of the membrane environment allowed us to show a deep penetration of FR into the lipid bilayer illustrating a possible access mode of FR into the cell. Dynamic nuclear polarization (DNP)-enhanced ssNMR was used to observe the structural response of specific segments of the Gα subunit to inhibitor binding. This revealed rigidification of the switch I binding site and an allosteric response in the α5 helix as well as suppression of structural changes induced by nucleotide exchange due to inhibition by FR. Our NMR studies of the FR-G protein complex conducted directly within a native membrane environment provide important insights into the inhibitors access via the lipid membrane, binding mode, and structural allosteric effects.
Competing Interests: There are no conflicts to declare.
(This journal is © The Royal Society of Chemistry.)
Databáze: MEDLINE
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