title: S-layer-streptavidin fusion proteins as template for nanopatterned molecular arrays.
abstract: Biomolecular self-assembly can be used as a powerful tool for nanoscale engineering. In this paper, we describe the development of building blocks for nanobiotechnology, which are based on the fusion of streptavidin to a crystalline bacterial cell surface layer (S-layer) protein with the inherent ability to self-assemble into a monomolecular protein lattice. The fusion proteins and streptavidin were produced independently in Escherichia coli, isolated, and mixed to refold and purify heterotetramers of 1:3 stoichiometry. Self-assembled chimeric S-layers could be formed in suspension, on liposomes, on silicon wafers, and on accessory cell wall polymer containing cell wall fragments. The two-dimensional protein crystals displayed streptavidin in defined repetitive spacing, and they were capable of binding d-biotin and biotinylated proteins. Therefore, the chimeric S-layer can be used as a self-assembling nanopatterned molecular affinity matrix to arrange biotinylated compounds on a surface. In addition, it has application potential as a functional coat of liposomes.
authors.affiliation: Center for Ultrastructure Research and Ludwig Boltzmann Institute for Molecular Nanotechnology, University of Agricultural Sciences, A-1180 Vienna, Austria.
authors.forename: Dieter
authors.initials: D
authors.lastname: Moll
authors.forename: Carina
authors.initials: C
authors.lastname: Huber
authors.forename: Birgit
authors.initials: B
authors.lastname: Schlegel
authors.forename: Dietmar
authors.initials: D
authors.lastname: Pum
authors.forename: Uwe B
authors.initials: UB
authors.lastname: Sleytr
authors.forename: Margit
authors.initials: M
authors.lastname: Sára
mesh_terms: D019921:Biotinylation; D007091:Image Processing, Computer-Assisted; D007700:Kinetics; D008081:Liposomes; D018625:Microscopy, Atomic Force / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D010539:Permeability; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism; D040081:Protein Array Analysis; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D019809:Streptavidin; D013698:Templates, Genetic
chemical_list: D008081:Liposomes; D011993:Recombinant Fusion Proteins; D019809:Streptavidin; D010544:Peroxidases