title: Supramolecularly oriented immobilization of proteins using cucurbit[8]uril.
abstract: A supramolecular strategy is used for oriented positioning of proteins on surfaces. A viologen-based guest molecule is attached to the surface, while a naphthol guest moiety is chemoselectively ligated to a yellow fluorescent protein. Cucurbit[8]uril (CB[8]) is used to link the proteins onto surfaces through specific charge-transfer interactions between naphthol and viologen inside the CB cavity. The assembly process is characterized using fluorescence and atomic force microscopy, surface plasmon resonance, IR-reflective absorption, and X-ray photoelectron spectroscopy measurements. Two different immobilization routes are followed to form patterns of the protein ternary complexes on the surfaces. Each immobilization route consists of three steps: (i) attaching the viologen to the glass using microcontact chemistry, (ii) blocking, and (iii) either incubation or microcontact printing of CB[8] and naphthol guests. In both cases uniform and stable fluorescent patterns are fabricated with a high signal-to-noise ratio. Control experiments confirm that CB[8] serves as a selective linking unit to form stable and homogeneous ternary surface-bound complexes as envisioned. The attachment of the yellow fluorescent protein complexes is shown to be reversible and reusable for assembly as studied using fluorescence microscopy.
authors.affiliation: Molecular Nanofabrication Group, Department of Science and Technology, MESA+ Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE, Enschede, The Netherlands.
authors.forename: Arántzazu
authors.initials: A
authors.lastname: González-Campo
authors.forename: Melanie
authors.initials: M
authors.lastname: Brasch
authors.forename: Dana A
authors.initials: DA
authors.lastname: Uhlenheuer
authors.forename: Alberto
authors.initials: A
authors.lastname: Gómez-Casado
authors.forename: Lanti
authors.initials: L
authors.lastname: Yang
authors.forename: Luc
authors.initials: L
authors.lastname: Brunsveld
authors.forename: Jurriaan
authors.initials: J
authors.lastname: Huskens
authors.forename: Pascal
authors.initials: P
authors.lastname: Jonkheijm
mesh_terms: D001665:Binding Sites; D001952:Bridged-Ring Compounds / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry; D008958:Models, Molecular; D009284:Naphthols / Q000737:chemistry; D011487:Protein Conformation; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013499:Surface Properties; D047028:Macrocyclic Compounds; D000099246:Cucurbit(n)urils
chemical_list: D001952:Bridged-Ring Compounds; D007093:Imidazoles; D055767:Immobilized Proteins; D008164:Luminescent Proteins; D009284:Naphthols; D000644:Quaternary Ammonium Compounds; C507198:cucurbit(8)uril; D047028:Macrocyclic Compounds; D000099246:Cucurbit(n)urils