title: Smart polymer brush nanostructures guide the self-assembly of pore-spanning lipid bilayers with integrated membrane proteins.
abstract: Nanopores in arrays on silicon chips are functionalized with pH-responsive poly(methacrylic acid) (PMAA) brushes and used as supports for pore-spanning lipid bilayers with integrated membrane proteins. Robust platforms are created by the covalent grafting of polymer brushes using surface-initiated atom transfer radical polymerization (ATRP), resulting in sensor chips that can be successfully reused over several assays. His-tagged proteins are selectively and reversibly bound to the nitrilotriacetic acid (NTA) functionalization of the PMAA brush, and consequently lipid bilayer membranes are formed. The enhanced membrane resistance as determined by electrochemical impedance spectroscopy and free diffusion of dyed lipids observed as fluorescence recovery after photobleaching confirmed the presence of lipid bilayers. Immobilization of the His-tagged membrane proteins on the NTA-modified PMAA brush near the pore edges is characterized by fluorescence microscopy. This system allows us to adjust the protein density in free-standing bilayers, which are stabilized by the polymer brush underneath. The potential application of the integrated platform for ion channel protein assays is demonstrated.
authors.affiliation: Materials Science and Technology of Polymers, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands. g.j.vancso@utwente.nl.
authors.forename: G Wilhelmina
authors.initials: GW
authors.lastname: de Groot
authors.forename: Sophie
authors.initials: S
authors.lastname: Demarche
authors.forename: M Gabriella
authors.initials: MG
authors.lastname: Santonicola
authors.forename: Louis
authors.initials: L
authors.lastname: Tiefenauer
authors.forename: G Julius
authors.initials: GJ
authors.lastname: Vancso
mesh_terms: D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000138:chemical synthesis / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D011109:Polymethacrylic Acids / Q000737:chemistry*
chemical_list: D008051:Lipid Bilayers; D008565:Membrane Proteins; D011109:Polymethacrylic Acids; C030613:polymethacrylic acid; D009571:Nitrilotriacetic Acid