A colorimetric fluorescent chemodosimeter based on diketopyrrolopyrrole and 1,3-indanedione for cysteine detection and cellular imaging in living cells (original) (raw)
Related papers
A cysteine-selective fluorescent probe for the cellular detection of cysteine
Biomaterials, 2012
A series of coumarin fluorophores (1e3), each bearing a double bond conjugated quinoline unit that can undergo a Michael-type reaction with thiol-containing compounds, is reported. These systems, designed to provide so-called turn-on changes in fluorescence response when exposed to thiols, act as fluorescent chemical sensors for cysteine (Cys), homocysteine (Hcy), and glutathione (GSH). In the case of 1, selectivity for Cys over Hcy and GSH is observed, both in terms of analyte-induced signal enhancement and response time. On the basis of fluorescence spectroscopic analyses, DFT calculations, and pH dependent studies this substrate selectivity is ascribed to steric interactions between the substituents on the quinolone units present in 1 and the targeted thiols, as well as to the comparatively lower pK a value of Cys relative to Hcy and GSH. In aqueous solution, probe 1 was found capable of detecting Cys with a detection limit of 10 À7 M. This system was successfully applied to the fluorescence imaging of intracellular Cys in HepG2 cells.
Chemical Science, 2012
General Information: Unless otherwise noted, materials were obtained from Aldrich and were used without further purification. 1 H NMR and 13 C NMR spectra were recorded on Brucker AM-300 spectrometers. 1 H NMR and 13 C NMR in CDCl 3 were measured on a Bruker AM-300 spectrometer with tetramethylsilane (TMS) as internal standard. Mass spectra were obtained using a JMS-HX 110A/110A Tandem Mass Spectrometer (JEOL). UV−vis spectra were obtained using a Scinco 3000 spectrophotometer (1 cm quartz cell) at 25 °C. Fluorescence spectra were recorded on RF-5301/PC (Shimada) fluorescence spectrophotometer (1 cm quartz cell) at 25 °C. Deionized water was used to prepare all aqueous solutions. Methods for cell culture and fluorescent imaging Human breast carcinoma (MCF-7) cells were seeded on 18 mm-glass coverslips (Marienfeld, Lauda-Koenigshofen, Germany) at density 2×10 5 cells and cultured in McCoy's 5a media with 10% bovine calf serum and 26 mM sodium carbonate at 37 ºC in a humidified incubator containing 5% CO 2 and 95% air. In order to induce oxidative stress, cells were rinsed three times with phosphate-buffered saline (PBS) and incubated in glucose-free DMEM (Dulbecco's Modified Eagle Media) without antibiotics and bovine calf serum for 2 h. 17 After the incubation, MCF-7 cells were rinsed with PBS and then incubated with 5 M of CyAc for 30 min at RT. The treated cells were washed with PBS and mounted onto a glass slide with ClearMount aqueous mounting medium (Invitrogen). To visualize the NIR fluorescence a zenon lamp (Hamamatsu, Shizuoka, Japan; 75 watt) and cy7 filter cube (Semrock, Rochester, NY; Ex. 660-750 nm/Em. 760-855 nm) was used in comparison with Hg 2+ lamp (Nikon; 100 watt) and Nikon filter cube (G-2A; Ex. 510-560 nm/Em. 590 nm) for 535 nm absorption peak of CyAc. Fluorescent images of the mounted cells were obtained by using an inverted microscope (Nikon Eclipse TE2000-U) at various magnifications (100 to 200 ).
Tetrahedron Letters, 2011
Two fluorescence probes for the detection of cysteine (Cys), glutathione (GSH) and other biothiols, such as homocysteine (Hcy) and cysteinyl-glycine (Cys-Gly), were developed. These molecular probes are coumarin-based derivatives containing a chalcone-like moiety that reacts with biothiols through a Michael addition reaction, leading to strong fluorescence enhancements. The reactivity of the tested biothiols toward both probes (ChC1 and ChC2) follows the order Cys > GSH > Hcy > Cys-Gly, ChC1 being less reactive than ChC2. Possible interference with other amino acids was assessed. ChC1 and ChC2 display a highly selective fluorescence enhancement with thiols, allowing these probes to be used for fluorimetric thiol determination in SH-SY5Y cells.
2021
A novel near-infrared fluorescent probe, namely propane-2,2-diylbis(2-((E)-2-(benzo[d]thiazol-2-yl)-2-cyanovinyl)-4,1-phenylene) diacrylate (BTA), was synthesized for selective detection of cysteine over other biologically significant amino acids. Upon addition of cysteine, the probe BTA displays a dramatic increase in fluorescence intensity at 715 nm along with a fast response time (4 min). The limit of detection (LOD) was calculated as 0.12 μM. In addition, the synthesized probe BTA was effectively utilized for the recognition of cysteine in blood serum and living cells.
Sensors and Actuators B: Chemical, 2014
NBD-chloride is widely used as an efficient probe for selective labelling of thiols in proteins due to formation sulfur-substituted NBD under physiological conditions. Selective conjugation involving thiolate group is favoured over amines of proteins because, amino-substituted NBD can be formed only under more basic and elevated temperature conditions. Sulfur-substituted NBDs generally display weak fluorescence properties compared to amino-substituted derivatives. However, a sulfur-substituted NBD can be converted to corresponding amino-substituted derivative via S-N Smiles rearrangement. Theoretical calculations predicted off-fluorescence state for either the probe or the sulfur-substituted NBD formed upon addition of cysteine. On-fluorescence state was predicted for corresponding amino-substituted NBD derivative. Based on UV-vis and fluorescence spectroscopic studies, most efficient rearrangement was observed for cysteine. The rearrangement was relatively slower for homocysteine and not feasible for glutathione. Detection of cysteine and homocysteine by the probe resulted in 1599-and 760-fold off-on fluorescence enhancements, respectively. Sensing of cysteine by the probe provided a detection limit of 2.0 × 10 −8 M. The sensing of intracellular cysteine by the probe was also demonstrated by live cell imaging.