Synthesis of functionalised fluorescent dyes and their coupling to amines and amino acids

Briggs, M. S. J.; Bruce, Ian J.; Miller, James N.; Moody, Christopher J.; Simmonds, A. C.; Swann, E.

doi:10.1039/a605012c

Cited by 64 publications

(57 citation statements)

References 5 publications

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“…6 Alkylation of these compounds with trimethylsilylethyl protected bromobutyric acid was achieved at 80 °C. The protecting group of 8a and b was effectively removed with 12.5% trifluoroacetic acid in dichloromethane.…”

Section: Synthesismentioning

confidence: 99%

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Probing lipid- and drug-binding domains with fluorescent dyes

Black

Stanley

Filipp

et al. 2008

Bioorganic & Medicinal Chemistry

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A series of 2-and 3-OH Nile red dyes was prepared in order to generate water-soluble probes that could be used to probe lipid binding to proteins. Various substitutions in positions 2-/3-, 6-, and 7-shifted wavelengths while maintaining the environmental sensitivity of Nile red. In order to increase the solubility of the dyes in aqueous solutions, we attached butyric acid groups to the 2-or 3-OH position. In addition, phenothiazine dyes, which exhibited particularly long excitation properties, were synthesized and tested for the first time. All dyes showed Stoke's shifts of 70-100 nm and changes in excitation and emission of over 100 nm, depending on the hydrophobicity of the environment. Binding studies with bovine serum albumin and the non-specific lipid transfer protein SCP2 revealed emission changes of more than 30 nm upon binding to the protein and a five-fold increase in emission intensity. Titration of the dye-loaded proteins with various lipids or drugs replaced the dye and thereby reversed the shift in wavelength intensity. This allowed us to estimate the lipid binding affinity of the investigated proteins. For SCP2, isothermal calorimetry (ITC) data verified the titration experiments. NMR titration experiments of SCP2 with Nile red 2-O-butyric acid (1a) revealed that the dye is bound within the lipid binding pocket and competes with lipid ligands for this binding site. These results give valuable insight into lipid and drug transport by proteins outside and inside cells.

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Section: Synthesismentioning

confidence: 99%

“…Such derivatives have previously been prepared by Briggs and co-workers as well as our own laboratory as fluorescent fatty acid derivatives. [6][7][8] Here we describe the synthesis of novel phenoxazine and phenothiazine derivatives in the form of their butyric acid derivatives (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Probing lipid- and drug-binding domains with fluorescent dyes

Black

Stanley

Filipp

et al. 2008

Bioorganic & Medicinal Chemistry

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“…16 First, the UV-vis signaling behavior of 1 toward representative alkali, alkaline earth, and transition metal ions was investigated. Compound 1 revealed a strong absorption at 580 nm in acetate buffered aqueous 50% methanol solution at pH 4.7 ( Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…The 2-hydroxy derivative of Nile Red was prepared following a reported procedure. 16 All solvents were purchased from Aldrich Chemical Co. as 'spectroscopic grade'. 1 H NMR (600 MHz) and 13 C NMR (150 MHz) spectra were obtained on a Varian VNS NMR spectrometer and referenced to the residual solvent signal.…”

Section: Methodsmentioning

confidence: 99%

Selective Mercuration of 2-Hydroxy Nile Red and Its Application towards Chemodosimetric Hg²⁺-selective Signaling

Lee¹,

Choi²,

Yu³

et al. 2010

Bulletin of the Korean Chemical Society

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Selective mercuration at the 1,6-positions of 2-hydroxy derivative of Nile Red and its application towards Hg 2+ -selective signaling was investigated. The 2-hydroxy Nile Red exhibited a selective UV-vis absorption and fluorescent signaling behavior towards Hg 2+ ions over common coexisting physiologically important metal ions in aqueous environment. 1H NMR studies revealed that the mercuration was selectively effected at the 1,6-positions of 2-hydroxy Nile Red, which is quite different from that at the 6,8-positions for the parent Nile Red.

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“…This Nile Red maleimide, with the thiolreactive maleimide unit spaced from the fluorescent Nile Red chromophore by a hexamethylene chain, was prepared from a Nile Red precursor bearing a phenolic hydroxyl group at the 2-position, 9-diethylamino-2-hydroxy-5H-benzo[a]phenoxazin-5-one. 40 The overall synthesis involves preparation of the phenolfunctionalized chromophore and a protected maleimide moiety followed by connection of the two parts via the C 6 spacer and finally deprotection to create the free maleimide. Full details of the synthetic method are presented in the Supplementary Material.…”

Section: Synthesis Of Nile Red Maleimide and Labeling Of Cys261mentioning

confidence: 99%

Probing the Sequence of Conformationally Induced Polarity Changes in the Molecular Chaperonin GroEL with Fluorescence Spectroscopy

et al. 2005

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Hydrophobic interactions play a major role in binding non-native substrate proteins in the central cavity of the bacterial chaperonin GroEL. The sequence of local conformational changes by which GroEL and its cofactor GroES assist protein folding can be explored using the polarity-sensitive fluorescence probe Nile Red. A specific single-cysteine mutant of GroEL (Cys261), whose cysteine is located inside the central cavity at the apical region of the protein, was covalently labeled with synthetically prepared Nile Red maleimide (NR). Bulk fluorescence spectra of Cys261-NR were measured to examine the effects of binding of the stringent substrate, malate dehydrogenase (MDH), GroES, and nucleotide on the local environment of the probe. After binding denatured substrate, the fluorescence intensity increased by 32±7%, suggesting enhanced hydrophobicity at the position of the label. On the other hand, in the presence of ATP, the fluorescence intensity decreased by 13±3%, implying increased local polarity. In order to explore the sequence of local polarity changes, substrate, GroES, and various nucleotides were added in different orders; the resulting changes in emission intensity provide insight into the sequence of conformational changes occurring during GroELmediated protein folding.

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Synthesis of functionalised fluorescent dyes and their coupling to amines and amino acids

Cited by 64 publications

References 5 publications

Probing lipid- and drug-binding domains with fluorescent dyes

Probing lipid- and drug-binding domains with fluorescent dyes

Selective Mercuration of 2-Hydroxy Nile Red and Its Application towards Chemodosimetric Hg²⁺-selective Signaling

Probing the Sequence of Conformationally Induced Polarity Changes in the Molecular Chaperonin GroEL with Fluorescence Spectroscopy

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Synthesis of functionalised fluorescent dyes and their coupling to amines and amino acids

Cited by 64 publications

References 5 publications

Probing lipid- and drug-binding domains with fluorescent dyes

Probing lipid- and drug-binding domains with fluorescent dyes

Selective Mercuration of 2-Hydroxy Nile Red and Its Application towards Chemodosimetric Hg2+-selective Signaling

Probing the Sequence of Conformationally Induced Polarity Changes in the Molecular Chaperonin GroEL with Fluorescence Spectroscopy

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Selective Mercuration of 2-Hydroxy Nile Red and Its Application towards Chemodosimetric Hg²⁺-selective Signaling