The Protein-Tethered Lipid Bilayer: A Novel Mimic of the Biological Membrane

• Published in: Biophysical journal Vol. 87; no. 5; pp. 3213 - 3220
• Main Authors: Giess, Frank, Friedrich, Marcel G., Heberle, Joachim, Naumann, Renate L., Knoll, Wolfgang
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.2004; Biophysical Society
• Subjects: Biomimetics - methods; Cell Membrane - chemistry; Cellular biology; Electric Impedance; Electron Transport Complex IV - chemistry; Histidine - chemistry; Lipid Bilayers - chemistry; Lipids; Macromolecular Substances; Membrane Fluidity; Membrane Proteins - chemistry; Membranes; Membranes, Artificial; Molecular Conformation; Protein Binding; Proteins
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1529/biophysj.104.046169

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 (C cO) 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.