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.

The Rylene Colorant Family—Tailored Nanoemitters for Photonics Research and Applications

“…The attachment of an NTA group within the imide structure of PDI (PDI-NTA 42; Figure 21) allows the site-specific labeling of proteins with an oligohistidine tag in the presence of nickel ions. [179] In contrast to other previously reported NTA-functionalized chromophores, the photophysical properties of the PDI-NTA remained unchanged upon complexation with nickel ions. [181][182][183] Protein labeling with this NTA-functionalized perylene(dicarboximide) was successfully demonstrated by applying oligo-His-tagged ATP synthase.…”

“…[33,39] In addition, the imide structure allows the introduction of two different kinds of substituents and, therefore, a single reactive group for biolabeling could be attached at this position. [179,180] In this way, water-soluble PDI and TDI dyes with four sulfonic acid groups as well as a single group that allows bioconjugation, for example, carboxylic acid or amino groups, suitable for formation of an amide bond with lysine, glutamic acid, or aspartic acid groups were obtained. [180] Chromophores 38-40 are highly soluble in water and have high fluorescence quantum yield.…”

Targeting cytochrome C oxidase in mitochondria with Pt(II)-porphyrins for photodynamic therapy

“…For confocal single-molecule imaging and localization in living cells, bright fluorophores with exceptional photo stability are required 19 . Thus new ultrastable dyes and fluorescent nanocrystals are continuously developed [20][21][22][23] . These fluorophores emit bursts of photons, which yield diffusion properties, association status and conformational changes in vitro and in vivo by fluorescence correlation spectroscopy [24][25][26][27][28] .…”

Novel derivatives of 1,6,7,12-tetrachloroperylene-3,4,9,10-tetracarboxylic acid: synthesis, electrochemical and optical properties

“…18,19,20,21 These dyes have also been used in other photo-physical processes of current interest, such as photocatalysis, 22,23 singlet exciton fission, 24,25 triplet-triplet annihilation, 26 organic photovoltaics, 27,28,29 lasing, 30 fluorescence probing, 31,32,33 and bio-labelling. 34 As compared to other organic dyes, perylene dyes provide a wealth of opportunities for the tuning of their properties via structural modification. These structural modifications can be achieved at three different positions around the perylene core; namely, the "peri" (3,4,9,10) positions, the "bay" (1, 6,7,12) positions, and the "ortho" (2,5,8,11) positions (Figure 1).…”