title: Reversible oriented surface immobilization of functional proteins on oxide surfaces.
abstract: Reversible and oriented immobilization of proteins in a functionally active form on solid surfaces is a prerequisite for the investigation of molecular interactions by surface-sensitive techniques. We demonstrate a method generally applicable for the attachment of proteins to oxide surfaces. A nitrilotriacetic acid group serving as a chelator for transition metal ions was covalently bound to the surface via silane chemistry. Reversible binding of the green fluorescent protein, modified with a hexahistidine extension, was monitored in situ using total internal reflection fluorescence. The association constant and kinetic parameters of the binding process were determined. The reversible, directed immobilization of proteins on surfaces as described here opens new ways for structural investigation of proteins and receptor-ligand interactions.
authors.affiliation: Laboratoire de Chimie Physique des Polymères et Membranes, Ecole Polytechnique Fédérale de Lausanne, Switzerland.
authors.forename: E L
authors.initials: EL
authors.lastname: Schmid
authors.forename: T A
authors.initials: TA
authors.lastname: Keller
authors.forename: Z
authors.initials: Z
authors.lastname: Dienes
authors.forename: H
authors.initials: H
authors.lastname: Vogel
mesh_terms: D002614:Chelating Agents / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006639:Histidine / Q000737:chemistry; D007700:Kinetics; D008164:Luminescent Proteins / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D009571:Nitrilotriacetic Acid / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011869:Radioligand Assay; D012821:Silanes / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000378:metabolism; D013499:Surface Properties
chemical_list: D002614:Chelating Agents; D008164:Luminescent Proteins; D008670:Metals; D011506:Proteins; D012821:Silanes; D013438:Sulfhydryl Compounds; D049452:Green Fluorescent Proteins; D006639:Histidine; D009571:Nitrilotriacetic Acid