title: Exploiting the nitrilotriacetic acid moiety for biolabeling with ultrastable perylene dyes.
abstract: Fluorescent probes are essential for the exploration of protein function, detection of molecular interactions, and conformational changes. The nitrilotriacetic acid derivatives of different chromophores were successfully used for site-selective noncovalent fluorescence labeling of histidine-tagged proteins. All of them, however, suffer from the same drawback--loss of the fluorescence upon binding of the nickel ions. Herein we present the solution and solid phase synthesis of water-soluble perylene(dicarboximide) functionalized with a nitrilotriacetic acid moiety (PDI-NTA). The photophysical properties of PDI-NTA revealed an exceptional photostability and fluorescence quantum yield that remained unchanged upon addition of nickel ions. The F1 complex of F0F1-ATP synthase from Escherichia coli, containing three hexahistidine tags, was labeled and the suitability for site-specific labeling of the new chromophore demonstrated using fluorescence correlation spectroscopy.
authors: Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
authors: Kalina
authors: K
authors: Peneva
authors: Gueorgui
authors: G
authors: Mihov
authors: Andreas
authors: A
authors: Herrmann
authors: Nawid
authors: N
authors: Zarrabi
authors: Michael
authors: M
authors: Börsch
authors: Thomas M
authors: TM
authors: Duncan
authors: Klaus
authors: K
authors: Müllen
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D001665:Binding Sites; D004926:Escherichia coli / Q000201:enzymology; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D009532:Nickel / Q000737:chemistry; D009571:Nitrilotriacetic Acid / Q000737:chemistry*; D010569:Perylene / Q000031:analogs & derivatives / Q000138:chemical synthesis*; D017785:Photons; D012995:Solubility; D014867:Water / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D014867:Water; D010569:Perylene; D009532:Nickel; D000251:Adenosine Triphosphatases; D009571:Nitrilotriacetic Acid