title: [Fluorescent fusion proteins with 10th human fibronectin domain].
abstract: In the current paper we describe a new type of hybrid molecules including red fluorescent protein mCherry and 10th type III human fibronectin domain (10Fn3) - one of the alternative scaffold proteins which can be used for the construction of antibody mimics with various binding specificity. We have constructed different gene variants encoding for the hybrid fluorescent protein and studied their expression in Escherichia coli cells. It was shown that N-terminal position of mCherry and modification of its N-terminal amino acid sequence promotes efficientbacterial expression of the hybrid protein in the soluble form. On the basis of the proposed construction we have obtained the hybrid fluorescent protein ChIBF, containing alphaVbeta3-integrin binding vari- ant of 10Fn3, and demonstrated the possibility of its utilization for the visualization of alphaVbeta3-integrin at the surface of MDCK epithelial cells by confocal microscopy.
authors.forename: L E
authors.initials: LE
authors.lastname: Petrovskaia
authors.forename: S Sh
authors.initials: SSh
authors.lastname: Gapizov
authors.forename: L N
authors.initials: LN
authors.lastname: Shingarova
authors.forename: E A
authors.initials: EA
authors.lastname: Kriukova
authors.forename: E F
authors.initials: EF
authors.lastname: Boldyreva
authors.forename: S A
authors.initials: SA
authors.lastname: Iakimov
authors.forename: E V
authors.initials: EV
authors.lastname: Svirshchevskaia
authors.forename: E P
authors.initials: EP
authors.lastname: Lukashev
authors.forename: D A
authors.initials: DA
authors.lastname: Dolgikh
authors.forename: M P
authors.initials: MP
authors.lastname: Kirpichnikov
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D004847:Epithelial Cells / Q000737:chemistry / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000276:immunology / Q000302:isolation & purification*; D008164:Luminescent Proteins / Q000737:chemistry*; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D000097573:Red Fluorescent Protein
chemical_list: D000906:Antibodies; D005353:Fibronectins; D039302:Integrin alphaVbeta3; D008164:Luminescent Proteins; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein