title: Supramolecular Protein Immobilization on Lipid Bilayers.
abstract: Protein immobilization on surfaces, and on lipid bilayers specifically, has great potential in biomolecular and biotechnological research. Of current special interest is the immobilization of proteins using supramolecular noncovalent interactions. This allows for a reversible immobilization and obviates the use of harsh ligation conditions that could denature fragile proteins. In the work presented here, reversible supramolecular immobilization of proteins on lipid bilayer surfaces was achieved by using the host-guest interaction of the macrocyclic molecule cucurbit[8]uril. A fluorescent protein was successfully immobilized on the lipid bilayer by making use of the property of cucurbit[8]uril to host together a methylviologen and the indole of a tryptophan positioned on the N-terminal of the protein. The supramolecular complex was anchored to the bilayer through a cholesterol moiety that was attached to the methylviologen tethered with a small polyethylene glycol spacer. Protein immobilization studies using a quartz crystal microbalance (QCM) showed the assembly of the supramolecular complexes on the bilayer. Specific immobilization through the protein N-terminus is more efficient than through protein side-chain events. Reversible surface release of the proteins could be achieved by washing with cucurbit[8]uril or buffer alone. The described system shows the potential of supramolecular assembly of proteins and provides a method for site-specific protein immobilization under mild conditions in a reversible manner.
authors.affiliation: Laboratory of Chemical Biology and Institute of Complex Molecular, Systems, Department of Biomedical Engineering, Eindhoven University of Technology, Den Dolech 2, 5612 AZ Eindhoven (The Netherlands).
authors.forename: Ralph P G
authors.initials: RP
authors.lastname: Bosmans
authors.affiliation: Advanced Soft Matter, Department of Chemical Engineering, Delft University of Technology, Julianalaan 136, 2628 BL Delft (The Netherlands).
authors.forename: Wouter E
authors.initials: WE
authors.lastname: Hendriksen
authors.affiliation: MESA+Institute for Nanotechnology and Department of Science and Technology, Laboratory Group of bioinspired molecular Engineering, University of Twente, Hallenweg 15, 7522 NB Enschede (The Netherlands).
authors.forename: Mark
authors.initials: M
authors.lastname: Verheijden
authors.affiliation: Advanced Soft Matter, Department of Chemical Engineering, Delft University of Technology, Julianalaan 136, 2628 BL Delft (The Netherlands).
authors.forename: Rienk
authors.initials: R
authors.lastname: Eelkema
authors.affiliation: MESA+Institute for Nanotechnology and Department of Science and Technology, Laboratory Group of bioinspired molecular Engineering, University of Twente, Hallenweg 15, 7522 NB Enschede (The Netherlands).
authors.forename: Pascal
authors.initials: P
authors.lastname: Jonkheijm
authors.affiliation: Advanced Soft Matter, Department of Chemical Engineering, Delft University of Technology, Julianalaan 136, 2628 BL Delft (The Netherlands).
authors.forename: Jan H
authors.initials: JH
authors.lastname: van Esch
authors.affiliation: Laboratory of Chemical Biology and Institute of Complex Molecular, Systems, Department of Biomedical Engineering, Eindhoven University of Technology, Den Dolech 2, 5612 AZ Eindhoven (The Netherlands). L.Brunsveld@tue.nl.
authors.forename: Luc
authors.initials: L
authors.lastname: Brunsveld
mesh_terms: D001952:Bridged-Ring Compounds / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D015394:Molecular Structure; D011092:Polyethylene Glycols / Q000737:chemistry*; D047028:Macrocyclic Compounds; D000099246:Cucurbit(n)urils
chemical_list: D001952:Bridged-Ring Compounds; D007093:Imidazoles; D055767:Immobilized Proteins; D008051:Lipid Bilayers; D011092:Polyethylene Glycols; C507198:cucurbit(8)uril; D047028:Macrocyclic Compounds; D000099246:Cucurbit(n)urils
keywords: immobilization; lipids; macrocycles; supramolecular chemistry; surface analysis