RESUMEN
Traditional biochemical methods for enzyme detection are mainly based on antibody-based immunoassays, which lack the ability to monitor the spatiotemporal distribution and, in particular, the in situ activity of enzymes in live cells and in vivo. In this review, we comprehensively summarize recent progress that has been made in the development of small-molecule as well as material-based fluorogenic probes for sensitive detection of the activities of enzymes that are related to a number of human diseases. The principles utilized to design these probes as well as their applications are reviewed. Specific attention is given to fluorogenic probes that have been developed for analysis of the activities of enzymes including oxidases and reductases, those that act on biomacromolecules including DNAs, proteins/peptides/amino acids, carbohydrates and lipids, and those that are responsible for translational modifications. We envision that this review will serve as an ideal reference for practitioners as well as beginners in relevant research fields.
Asunto(s)
Enzimas/química , Colorantes Fluorescentes/química , Animales , Biomarcadores/metabolismo , Enzimas/metabolismo , Glicósido Hidrolasas/química , Glicósido Hidrolasas/metabolismo , Humanos , Neoplasias/diagnóstico , Neoplasias/enzimología , Enfermedades Neurodegenerativas/diagnóstico , Enfermedades Neurodegenerativas/enzimología , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Péptido Hidrolasas/química , Péptido Hidrolasas/metabolismoRESUMEN
A mitochondria-targeting ratiometric probe was designed for γ-glutamyltranspeptidase (γGT). Mechanistic study by HPLC and an inhibitor assay showed that the probe underwent γGT-mediated amide-to-amine transformation and induced a ratiometric fluorescence response in cellular mitochondria. Further application was successful for the detection of cancerous colons in mice.
Asunto(s)
Neoplasias del Colon/diagnóstico por imagen , Colorantes Fluorescentes/química , Glutamina/análogos & derivados , Glutamina/química , Indoles/química , Mitocondrias/metabolismo , gamma-Glutamiltransferasa/análisis , Animales , Fluorescencia , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/metabolismo , Glutamina/síntesis química , Glutamina/metabolismo , Células HCT116 , Humanos , Hidrólisis , Indoles/síntesis química , Indoles/metabolismo , Límite de Detección , Ratones , gamma-Glutamiltransferasa/metabolismoRESUMEN
We report a chromogenic and fluorescence turn-on probe based on crotonoyl ester-functionalized oxazolidinoindole for the selective detection of cysteine in neutral buffer. The probe rapidly formed indocyanophenolate through the Michael addition and a subsequent cyclization reaction of cysteine, inducing both a dramatic bathochromic shift (>130 nm) and a large fluorescence turn-on response (F/F0 12) in the UV-vis and fluorescence spectra and affording a micromolar limit of detection (LOD = 5.0 µM) of cysteine in HEPES buffer. When cysteine was added, the probe exhibited a dual optical change with strong green fluorescence and dramatic red color by the oxazolidinoindole-to-hydroxyethylindolium transformation. Further cellular application of the probe was successfully performed for the mitochondrial imaging of HeLa cells.
Asunto(s)
Cisteína/análisis , Colorantes Fluorescentes/química , Indoles/química , Mitocondrias/metabolismo , Oxazoles/química , Crotonatos/química , Fluorescencia , Colorantes Fluorescentes/síntesis química , Células HeLa , Humanos , Indoles/síntesis química , Límite de Detección , Microscopía Confocal/métodos , Microscopía Fluorescente/métodos , Modelos Químicos , Oxazoles/síntesis químicaRESUMEN
Homocysteine and methylmalonic acid are important biomarkers for diseases associated with an impaired central nervous system (CNS). A new chemoassay utilizing coumarin-based fluorescent probe 1 to detect the levels of homocysteine is successfully implemented using Parkinson's disease (PD) patients' blood serum. In addition, a rapid identification of homocysteine and methylmalonic acid levels in blood serum of PD patients was also performed using the liquid chromatography-mass spectrometry (LC-MS). The results obtained from both analyses were in agreement. The new chemoassay utilizing coumarin-based fluorescent probe 1 offers a cost- and time-effective method to identify the biomarkers in CNS patients.
Asunto(s)
Cumarinas , Colorantes Fluorescentes , Homocisteína/sangre , Ácido Metilmalónico/sangre , Enfermedad de Parkinson/sangre , Enfermedad de Parkinson/diagnóstico , Espectrometría de Fluorescencia/métodos , Biomarcadores/sangre , Técnicas Biosensibles/métodos , Humanos , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Although a lot of mitochondria-targeting biothiol probes have been developed and applied to cellular imaging through thiol-induced disulfide cleavage or Michael addition reactions, relatively few probes assess mitochondrial GSH with high selectivity over Cys and Hcy and with NIR fluorescence capable of noninvasive imaging in biological samples. In order to monitor mitochondrial GSH with low background autofluorescence, we designed a heptamethine-azo conjugate as an NIR fluorescent probe by introducing a tunable lipophilic cation unit as the biomarker for mitochondria and a nitroazo group as the GSH-selective reaction site as well as the fluorescence quencher. The probe exhibited a dramatic off-on NIR fluorescence response toward GSH with high selectivity over other amino acids including Cys and Hcy. Further application to cellular imaging indicated that the probe was highly responsive to the changes of mitochondrial GSH in cells.
Asunto(s)
Compuestos Azo/química , Cisteína/análisis , Colorantes Fluorescentes/química , Glutatión/análisis , Homocisteína/análisis , Indoles/química , Células HeLa , Humanos , Microscopía Confocal , Mitocondrias/química , Imagen Óptica , Espectrometría de FluorescenciaRESUMEN
We developed a fluorescein aldehyde probe with disulfide functionality for the fluorescence detection of biologically important thiols. The probe displayed highly selective responses to cysteine (Cys) and homocysteine (Hcy) over glutathione (GSH) due to the rapid ring formation reaction of Cys and Hcy with the aldehyde group of the probe and the concomitant cleavage of the disulfide group followed by subsequent intramolecular cyclization. The fluorescent probe also exhibited a highly sensitive fluorescence turn-on response to Hcy with a detection limit of 2.4 µM Hcy in HEPES buffer.
Asunto(s)
Aldehídos/química , Cisteína/análisis , Disulfuros/química , Fluoresceína/química , Colorantes Fluorescentes/química , Homocisteína/análisis , Tampones (Química) , Cisteína/química , Colorantes Fluorescentes/análisis , Colorantes Fluorescentes/síntesis química , HEPES/química , Homocisteína/química , Estructura MolecularRESUMEN
A fluorescent probe (1) with a hydrogen bond was designed for the detection of GSH. The probe exhibited a rapid and ratiometric fluorescence response to GSH through a Michael reaction and allowed us to obtain clear cellular images for GSH.
Asunto(s)
Cumarinas/síntesis química , Colorantes Fluorescentes/síntesis química , Glutatión/análisis , Cumarinas/química , Fluorescencia , Colorantes Fluorescentes/química , Células HeLa , Humanos , Enlace de Hidrógeno , Estructura MolecularRESUMEN
A nonfluorescent coumarin-malonitrile conjugate (1) was transformed into a strongly fluorescent molecule through the Michael addition of a thiol group to the α,ß-unsaturated malonitrile of 1. The molecular probe has exhibited a highly selective fluorescence response toward biothiols (Cys, Hcy, GSH) with micromolar sensitivity both in vitro and in vivo.
Asunto(s)
Cumarinas/química , Colorantes Fluorescentes/química , Sondas Moleculares/química , Nitrilos/química , Espectrometría de Fluorescencia/métodos , Compuestos de Sulfhidrilo/análisis , Cisteína/análisis , Cisteína/química , Fluorescencia , Glutatión/análisis , Glutatión/química , Homocisteína/análisis , Homocisteína/química , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Compuestos de Sulfhidrilo/química , Factores de TiempoRESUMEN
An activated Michael acceptor type of probe by an intramolecular hydrogen bond has shown a selective fluorescence turn-on response to cyanide through a conjugated addition of the nucleophilic anion to the enone probe with a 1300-fold increase in its fluorescence intensity.
Asunto(s)
Fenómenos Químicos , Cianuros/análisis , Colorantes Fluorescentes , Enlace de Hidrógeno , Sitios de Unión , FluorescenciaRESUMEN
A latent fluorophore (1) was rationally designed to give rise to strong fluorescence through a selective gold ion-mediated hydroarylation reaction by a catalytic amount of the gold(III) ion. It is a highly selective and sensitive fluorescence turn-on probe for gold(III) ions and is operative even in protic solvents.
RESUMEN
A Co(II)-salen based fluorescent sensor (1.Co) that can selectively recognize cyanide anions in 1:2 binding stoichiometry over other anions has been developed. 1.Co displayed fluorescence enhancement upon the addition of cyanide owing to the interruption of photoinduced electron transfer from the coumarin fluorophore to the cobalt(II) ion. A general regression method was developed to calculate the binding constants in the 1:2 binding system, through which the 1:2 binding between 1.Co and cyanide anions was estimated to be in the range of micromolar dissociation constants.
Asunto(s)
Cobalto/química , Cumarinas/síntesis química , Colorantes Fluorescentes/síntesis química , Nitrilos/análisis , Compuestos Organometálicos/síntesis química , Aniones , Cumarinas/química , Fluorescencia , Colorantes Fluorescentes/química , Estructura Molecular , Compuestos Organometálicos/química , Espectrometría de FluorescenciaRESUMEN
The first "Off-On" type fluorescent probe for Au3+ is reported, in which rhodamine-alkyne derivative displayed highly selective fluorescence enhancement and "naked-eye" detection upon the addition of Au3+ at pH 7.4.
Asunto(s)
Oro/análisis , Pargilina/análogos & derivados , Propilaminas/química , Alquinos , Cationes , Colorimetría , Ciclización , Colorantes Fluorescentes , Pargilina/química , RodaminasRESUMEN
Hammett plot reveals that there is a significant electronic effect on the rate of resonance assisted hydrogen bond (RAHB) directed diaza-Cope rearrangement reaction with a rho value of 1.6. DFT computation shows that the rearrangement reaction becomes thermodynamically more favorable for the substrates with electron withdrawing substituents. A substrate with the nitro substituent (1a) reacts about 50-fold faster than that with the methoxy substituent (1g).
RESUMEN
A simple fluorescent probe based on an ortho-hydroxy aldehyde-functionalized coumarin showed selective responses to homocysteine and cysteine by fluorescence turn-on.
Asunto(s)
Cisteína/análisis , Cisteína/química , Colorantes Fluorescentes/análisis , Colorantes Fluorescentes/química , Homocisteína/análisis , Homocisteína/química , Agua/química , Espectroscopía de Resonancia Magnética , Estructura Molecular , Espectrometría de FluorescenciaRESUMEN
A dinuclear copper complex that binds tightly and selectively to oxalate over other dicarboxylates like malonate, succinate, and glutarate has been developed. This receptor can be used for fluorescent detection of oxalate in water at physicological pH by chemosensing ensemble approach. Crystal structure of oxalate bound to the receptor together with molecular mechanics and DFT computations provide insights into the tight and selective binding of the anion by the receptor.
Asunto(s)
Cobre/química , Oxalatos/análisis , Agua/análisis , Cristalografía por Rayos X , Eosina Amarillenta-(YS)/química , Colorantes Fluorescentes/química , Concentración de Iones de Hidrógeno , Naftalenos/química , Compuestos Organometálicos/química , Oxalatos/química , Espectrometría de Fluorescencia/métodosRESUMEN
A coumarin-based fluorescent chemodosimeter with a salicylaldehyde functionality as a binding site has been developed for selective detection of cyanide anions over other anions in water at biological pH.
Asunto(s)
Cumarinas/síntesis química , Cianuros/análisis , Colorantes Fluorescentes/síntesis química , Abastecimiento de Agua/análisis , Aldehídos/química , Cumarinas/química , Colorantes Fluorescentes/química , Concentración de Iones de Hidrógeno , Estructura Molecular , Espectrometría de Fluorescencia/métodos , EspectrofotometríaRESUMEN
A cobalt(III)-salen complex (3) with an axial substituent on the diamine backbone has been synthesized. Crystal structure reveals that the axial substituent (p-nitrophenyl group) is positioned in close proximity to the metal binding site. The stereoselectivity of the cobalt complex for binding amino alcohols increases with increasing steric bulk of the amino alcohol from alaninol (2.9) to valinol (6.2) and t-leucinol (36.0).
RESUMEN
Crystal structure and activation entropy data indicate that H-bond directed diaza-Cope rearrangement of chiral diimines takes place with a high degree of preorganization. CD spectroscopy and HPLC data show that there is inversion of stereochemistry for the reaction with excellent enantioselectivity.
RESUMEN
A chiral aldehyde with three H-bond donating groups (2) has been synthesized. This aldehyde binds a variety of chiral 1,2-amino alcohols in benzene with the same sense of stereoselectivity. Computational and experimental data indicate that one imine bond, one resonance-assisted H-bond to the imine nitrogen, and two H-bonds to the alcoholic oxygen all play an important role in the stereoselective recognition. [structure: see text]