title: EGFR Dynamics Change during Activation in Native Membranes as Revealed by NMR.
abstract: The epidermal growth factor receptor (EGFR) represents one of the most common target proteins in anti-cancer therapy. To directly examine the structural and dynamical properties of EGFR activation by the epidermal growth factor (EGF) in native membranes, we have developed a solid-state nuclear magnetic resonance (ssNMR)-based approach supported by dynamic nuclear polarization (DNP). In contrast to previous crystallographic results, our experiments show that the ligand-free state of the extracellular domain (ECD) is highly dynamic, while the intracellular kinase domain (KD) is rigid. Ligand binding restricts the overall and local motion of EGFR domains, including the ECD and the C-terminal region. We propose that the reduction in conformational entropy of the ECD by ligand binding favors the cooperative binding required for receptor dimerization, causing allosteric activation of the intracellular tyrosine kinase.
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Kaplan
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Siddarth
authors.initials: S
authors.lastname: Narasimhan
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Cecilia
authors.initials: C
authors.lastname: de Heus
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Deni
authors.initials: D
authors.lastname: Mance
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: van Doorn
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Klaartje
authors.initials: K
authors.lastname: Houben
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Dušan
authors.initials: D
authors.lastname: Popov-Čeleketić
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Reinier
authors.initials: R
authors.lastname: Damman
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Katrukha
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Purvi
authors.initials: P
authors.lastname: Jain
authors.affiliation: Biomolecular Imaging, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Willie J C
authors.initials: WJC
authors.lastname: Geerts
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Albert J R
authors.initials: AJR
authors.lastname: Heck
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Gert E
authors.initials: GE
authors.lastname: Folkers
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Lukas C
authors.initials: LC
authors.lastname: Kapitein
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Simone
authors.initials: S
authors.lastname: Lemeer
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands. Electronic address: p.vanbergen@uu.nl.
authors.forename: Paul M P
authors.initials: PMP
authors.lastname: van Bergen En Henegouwen
authors.affiliation: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, the Netherlands. Electronic address: m.baldus@uu.nl.
authors.forename: Marc
authors.initials: M
authors.lastname: Baldus
mesh_terms: D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D055503:Protein Multimerization; D013816:Thermodynamics; D022161:Transport Vesicles / Q000737:chemistry
chemical_list: D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: EGFR; NMR; Solid-state NMR; activation; cancer; membrane protein; receptor; tyrosine kinase