title: Reversible, site-specific immobilization of polyarginine-tagged fusion proteins on mica surfaces.
abstract: A large variety of genes is expressed as fusion proteins for the purpose of characterization and purification in molecular biology. We have used this strategy to append polyarginine peptides in order to achieve specific binding of the Arg-tag to atomically flat, negatively charged mica surfaces. We show that the model protein, hexaarginine-tagged green fluorescent protein (GFP), binds to mica via its Arg-tag based on ion exchange of naturally occurring potassium cations. Only non-specific binding was observed with the control protein that is free of the Arg-tag. This novel technology will be widely applicable to orient functional proteins on flat surfaces.
authors.affiliation: Department of Biochemistry, Beckman Center B405, Stanford University Medical Center, CA 94305-5307, USA.
authors.forename: S
authors.initials: S
authors.lastname: Nock
authors.forename: J A
authors.initials: JA
authors.lastname: Spudich
authors.forename: P
authors.initials: P
authors.lastname: Wagner
mesh_terms: D000327:Adsorption; D000538:Aluminum Silicates; D000595:Amino Acid Sequence; D001120:Arginine; D001483:Base Sequence; D001665:Binding Sites; D049452:Green Fluorescent Proteins; D006639:Histidine; D008164:Luminescent Proteins / Q000737:chemistry* / Q000302:isolation & purification; D008239:Lysine; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D016324:Sequence Tagged Sites
chemical_list: D000538:Aluminum Silicates; D008164:Luminescent Proteins; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C015462:polyarginine; C033223:polyhistidine; D006639:Histidine; D001120:Arginine; D008239:Lysine; C011934:mica