Exploiting the nitrilotriacetic acid moiety for biolabeling with ultrastable perylene dyes.
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.
Peneva
Kalina
K
Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
Mihov
Gueorgui
G
Herrmann
Andreas
A
Zarrabi
Nawid
N
Börsch
Michael
M
Duncan
Thomas M
TM
Müllen
Klaus
K
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
D005456:Fluorescent Dyes; D014867:Water; D010569:Perylene; D009532:Nickel; D000251:Adenosine Triphosphatases; D009571:Nitrilotriacetic Acid