title: N-Glycosylation as determinant of epidermal growth factor receptor conformation in membranes.
abstract: The epidermal growth factor receptor (EGFR) regulates several critical cellular processes and is an important target for cancer therapy. In lieu of a crystallographic structure of the complete receptor, atomistic molecular dynamics (MD) simulations have recently shown that they can excel in studies of the full-length receptor. Here we present atomistic MD simulations of the monomeric N-glycosylated human EGFR in biomimetic lipid bilayers that are, in parallel, also used for the reconstitution of full-length receptors. This combination enabled us to experimentally validate our simulations, using ligand binding assays and antibodies to monitor the conformational properties of the receptor reconstituted into membranes. We find that N-glycosylation is a critical determinant of EGFR conformation, and specifically the orientation of the EGFR ectodomain relative to the membrane. In the absence of a structure for full-length, posttranslationally modified membrane receptors, our approach offers new means to structurally define and experimentally validate functional properties of cell surface receptors in biomimetic membrane environments.
authors: Department of Physics, Tampere University of Technology, FI-33101 Tampere, Finland;
authors: Karol
authors: K
authors: Kaszuba
authors: Paul Langerhans Institute Dresden of the Helmholtz Centre Munich at the University Clinic Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany; German Center for Diabetes Research (DZD e.V.), 85764 Neuherberg, Germany;
authors: Michał
authors: M
authors: Grzybek
authors: Department of Physics, Tampere University of Technology, FI-33101 Tampere, Finland;
authors: Adam
authors: A
authors: Orłowski
authors: Department of Physics, Tampere University of Technology, FI-33101 Tampere, Finland;
authors: Reinis
authors: R
authors: Danne
authors: Department of Physics, Tampere University of Technology, FI-33101 Tampere, Finland;
authors: Tomasz
authors: T
authors: Róg
authors: Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany; and simons@mpi-cbg.de coskun@plid.de ilpo.vattulainen@tut.fi.
authors: Kai
authors: K
authors: Simons
authors: Paul Langerhans Institute Dresden of the Helmholtz Centre Munich at the University Clinic Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany; German Center for Diabetes Research (DZD e.V.), 85764 Neuherberg, Germany; simons@mpi-cbg.de coskun@plid.de ilpo.vattulainen@tut.fi.
authors: Ünal
authors: Ü
authors: Coskun
authors: Department of Physics, Tampere University of Technology, FI-33101 Tampere, Finland; Center for Biomembrane Physics (MEMPHYS), University of Southern Denmark, 5230 Odense, Denmark simons@mpi-cbg.de coskun@plid.de ilpo.vattulainen@tut.fi.
authors: Ilpo
authors: I
authors: Vattulainen
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D003198:Computer Simulation; D066246:ErbB Receptors / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011510:Proteolipids / Q000737:chemistry; D012984:Software
chemical_list: D000911:Antibodies, Monoclonal; D008024:Ligands; D008051:Lipid Bilayers; D011510:Proteolipids; C020485:proteoliposomes; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: EGFR; MD simulation; lipids; lipid–protein interaction; proteoliposomes