EGFR Dynamics Change during Activation in Native Membranes as Revealed by NMR
- Creators
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Kaplan, Mohammed
- Narasimhan, Siddarth
- de Heus, Cecilia
- Mance, Deni
- van Doorn, Sander
- Houben, Klaartje
- Popov-Čeleketić, Dušan
- Damman, Reinier
- Katrukha, Eugene A.
- Jain, Purvi
- Geerts, Willie J. C.
- Heck, Albert J. R.
- Folkers, Gert E.
- Kapitein, Lukas C.
- Lemeer, Simone
- van Bergen en Henegouwen, Paul M. P.
- Baldus, Marc
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.
Additional Information
© 2016 Elsevier. Received 4 May 2016; revised 8 August 2016; accepted 20 October 2016; available online 10 November 2016. We thank Willem Kegel and Markus Weingarth for helpful discussions and Johan van der Zwan for technical support. This work was funded in part by Netherlands Organization for Scientific Research (NWO) (grants 700.26.121 and 700.10.443 to M.B, STW12152 to P.B.H. and a VIDI grant 723.013.008 to SL) and iNEXT (project number 653706), a Horizon 2020 program of the European Union. In addition, S.v.D., S.L., and A.J.R.H. are supported by the project Proteins At Work (project 184.032.201), a program of the Netherlands Proteomics Centre financed by NWO as part of the National Roadmap Largescale Research Facilities of the Netherlands. The NMR experiments were supported in part by uNMR-NL, an NWO-funded National Roadmap Large-Scale Facility of the Netherlands. We are indebted to Paul Tordo and Olivier Ouari (Marseille) for providing AMUPol.Attached Files
Supplemental Material - 1-s2.0-S0092867416314593-supp.pdf
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Additional details
- Eprint ID
- 73191
- Resolver ID
- CaltechAUTHORS:20170103-160614195
- 700.26.121
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- 700.10.443
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- STW12152
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- 723.013.008
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- 653706
- iNEXT
- 184.032.201
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- National Ultrahigh-field Nuclear Magnetic Resonance Facility (uNMR-NL)
- Created
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2017-01-04Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field