title: The protein-tethered lipid bilayer: a novel mimic of the biological membrane.
abstract: A new concept of solid-supported tethered bilayer lipid membrane (tBLM) for the functional incorporation of membrane proteins is introduced. The incorporated protein itself acts as the tethering molecule resulting in a versatile system in which the protein determines the characteristics of the submembraneous space. This architecture is achieved through a metal chelating surface, to which histidine-tagged (His-tagged) membrane proteins are able to bind in a reversible manner. The tethered bilayer lipid membrane is generated by substitution of protein-bound detergent molecules with lipids using in-situ dialysis or adsorption. The system is characterized by surface plasmon resonance, quartz crystal microbalance, and electrochemical impedance spectroscopy. His-tagged cytochrome c oxidase (CcO) is used as a model protein in this study. However, the new system should be applicable to all recombinant membrane proteins bearing a terminal His-tag. In particular, combination of surface immobilization and membrane reconstitution opens new prospects for the investigation of functional membrane proteins by various surface-sensitive techniques under a defined electric field.
authors.affiliation: Max Planck Institute for Polymer Research, Mainz, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Giess
authors.forename: Marcel G
authors.initials: MG
authors.lastname: Friedrich
authors.forename: Joachim
authors.initials: J
authors.lastname: Heberle
authors.forename: Renate L
authors.initials: RL
authors.lastname: Naumann
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Knoll
mesh_terms: D032701:Biomimetics / Q000379:methods*; D002462:Cell Membrane / Q000737:chemistry*; D017097:Electric Impedance; D003576:Electron Transport Complex IV / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000737:chemistry*; D008567:Membranes, Artificial; D008968:Molecular Conformation; D011485:Protein Binding
chemical_list: D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008565:Membrane Proteins; D008567:Membranes, Artificial; D006639:Histidine; D003576:Electron Transport Complex IV