title: Generation of artificial channels by multimerization of β-strands from natural porin.
abstract: General diffusion porins are passive transmembrane channels. We have explored the possibility to create artificial nanopores starting from natural β-barrel structures. Structural elements of bacterial porins were used to build a series of artificial nanopores. The basic module was selected by multi-alignment of general diffusion porins. The sequence corresponded to a highly conserved motif containing two β-strands, which was obtained from Escherichia coli OmpF. Dimeric to octameric repeats were obtained through cDNA recombinant technology. The hexameric repeat was used to test its properties. This protein was expressed, purified and reconstituted in the planar bilayer membranes. It was able to form channels in membranes with a conductance of 300 pS in 150 mm KCl and did not show any relevant voltage-dependence.
authors.affiliation: Department of Biology, Laboratory of Biochemistry and Molecular Biology, University of Catania, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Lolicato
authors.forename: Simona
authors.initials: S
authors.lastname: Reina
authors.forename: Angela
authors.initials: A
authors.lastname: Messina
authors.forename: Francesca
authors.initials: F
authors.lastname: Guarino
authors.forename: Mathias
authors.initials: M
authors.lastname: Winterhalter
authors.forename: Roland
authors.initials: R
authors.lastname: Benz
authors.forename: Vito
authors.initials: V
authors.lastname: De Pinto
mesh_terms: D000595:Amino Acid Sequence; D019295:Computational Biology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D058608:Nanopores; D018272:Porins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: C109237:OmpF protein; D018272:Porins; D011994:Recombinant Proteins