title: Engineering a circularly permuted GFP scaffold for peptide presentation.
abstract: The use of peptides as in vivo and in vitro ligand binding agents is hampered by the high flexibility, low stability and lack of intrinsic detection signal of peptide aptamers. Recent attempts to overcome these limitations included the integration of the binding peptide into a stable protein scaffold. In this paper, we present the optimization and testing of a circularly permuted variant of the green fluorescent protein (GFP). We examined the ability of the optimized scaffold to accept peptide insertions at three different regions. The three regions chosen are localized in close spatial proximity to each other and support different conformations of the inserted peptides. In all the three regions peptides with a biased, but still comprehensive, amino acid repertoire could be presented without disturbing the function of the optimized GFP-scaffold.
authors.affiliation: Institut für Biochemie, Charité-Universitätsmedizin Berlin, Monbijoustrasse 20, D-10117 Berlin, Germany. mailto:paschke@jerini.de
authors.forename: Matthias
authors.initials: M
authors.lastname: Paschke
authors.forename: Christian
authors.initials: C
authors.lastname: Tiede
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Höhne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism*; D019791:Guanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011489:Protein Denaturation / Q000187:drug effects; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
chemical_list: D019151:Peptide Library; D010455:Peptides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA; D019791:Guanidine