RESUMEN
In search of new probes for biomolecules, the spectral fluorescent study of four monomethine cyanine dyes (MCD), both unsymmetrical and symmetrical, has been carried out in different organic solvents, in aqueous buffer solutions, and in the presence of DNA and HSA. The complexation of MCD with biomacromolecules leads to a steep growth of the fluorescence intensity. Complexes of MCD with dsDNA and HSA of various types were modeled in silico by molecular docking. Experiments on thermal dissociation of dsDNA in the presence of MCD showed the formation of intercalative complexes of MCD with DNA. Quenching of intrinsic fluorescence of HSA by MCD occurred with rate constants much higher than the diffusion limit, that is, in dye-HSA complexes. Effective constants of MCD complexation with the biomacromolecules were estimated. MCD 1 has the best characteristics as a possible fluorescent probe for dsDNA and can serve as a sensitive and selective probe for dsDNA in the presence of HSA. Photochemical properties of MCD complexed with DNA have been also studied. An increase in the quantum yield of the triplet states of MCD in complexes with DNA has been found, which may be important for using these dyes as potential candidates in photodynamic therapy.
Asunto(s)
Fotoquimioterapia , Quinolinas , Simulación del Acoplamiento Molecular , Colorantes Fluorescentes , DifusiónRESUMEN
Spectral-fluorescent properties of the meso-substituted anionic cyanine dye 3,3'-di-(γ-sulfopropyl)-4,5,4',5'-dibenzo-9-methylthiacarbocyanine betaine (DMC) were studied in solutions and in the presence of human serum albumin (HSA). The properties of DMC were compared with those of the previously studied meso-substituted anionic dyes 3,3'-di(γ-sulfopropyl)-4,5,4',5'-dibenzo-9-ethylthiacarbocyanine betaine (DEC), 3,3'-di-(γ-sulfopropyl)-9-methylthiacarbocyanine betaine (MTC) and 3,3'-di-(γ-sulfopropyl)-5,5'-diphenyl-9-ethyloxacarbocyanine betaine (OCC), which were studied here in more detail. In aqueous solutions, DMC, like DEC, is prone to dimerization; it also forms H- and J-aggregates. The noncovalent interaction of DMC with HSA leads to decomposition of the dimers with a shift in the cis-trans isomeric equilibrium toward the trans-monomer complexed with HSA, which is accompanied by a significant increase in fluorescence. The spectral-fluorescent data were used to estimate the binding constants of the dyes with HSA and other characteristics of the dyes, which are important when used as probes for HSA. The effect of structural rearrangements of HSA upon denaturation by urea on the spectral-fluorescent properties of the dyes was studied. Molecular docking of the dye-HSA systems was performed. A comparative assessment of the prospects for the use of the dyes as spectral-fluorescent probes for HSA in vitro was carried out.
Asunto(s)
Carbocianinas/química , Albúmina Sérica Humana/química , Aniones/química , Humanos , Simulación del Acoplamiento Molecular , Estructura MolecularRESUMEN
In this work, a novel and facile ratiometric fluorescence probe was prepared for the visual detection of dopamine (DA). In this detection system, red-emission CdTe@SiO2 (r-QDs@SiO2) was used as steady core of the probe and inverse microemulsion method was applied to synthesize uniform r-QDs@SiO2, this step could protect CdTe from contacting with human skin directly. Polydopamine (PDA) acted as response signal to detect DA, a very handy method which just combined polyethyleneimine (PEI) with DA together to synthesize PDA, this way for synthesis of PDA was much time-saving and non-toxic than any other methods. Differently from traditional analysis processes, the products of this experiment were also the analysis substances in final. Under optimum measurement conditions, the dual-emission ratiometric fluorescence probe was used for detections of DA in a concentration ranged from 10µM to 80µM with a detection limit of 0.12µM, with addition of DA the color of the probe changed from red to green watched by naked eyes. In addition, the developed probe was also used for detections of DA in human serum samples successfully. This study provides a simple, time-saving and non-toxic approach for detections of DA without the requirement of complex equipment.
Asunto(s)
Técnicas Biosensibles/métodos , Fluorescencia , Colorantes Fluorescentes/química , Indoles/sangre , Espectrometría de Fluorescencia/métodos , Humanos , Límite de Detección , PolímerosRESUMEN
The noncovalent interaction of the polymethine dye probe 3,3',9-trimethylthiacarbocyanine iodide (Cyan 2) with chondroitin-4-sulfate (C4S) in buffer solutions with different pH and in water in the absence of buffers has been studied by spectral-fluorescent methods. It has been shown that in all media studied, at relatively high concentrations, the dye is bound to C4S mainly as a monomer, which is accompanied by a steep rise of fluorescence (the intermediate formation of dye aggregates on the biopolymer is also observed). From the dependence of the fluorescence quantum yield on the concentration of C4S, the parameters of binding of the dye monomer to C4S were obtained: the effective binding constant K, the number of the monomeric C4S units n per one dye monomer bound to C4S, and the fluorescence quantum yield of the bound dye monomer Φfb. The dependence of Φfb (and K) on ÑÐ of the medium is not monotonic: it has a minimum in the region of neutral pH and a growth in the regions of acid and basic pH. This can be explained by changing the charge of a C4S macromolecule as a function of pH and related conformational alterations in the biopolymer, which can affect the rigidity of a dye molecule and the energy of its interaction with the biopolymer.