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1.
Chem Commun (Camb) ; 56(63): 9008-9011, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32638755

RESUMO

Herein, a novel strategy for in situ imaging and real-time monitoring of intracellular tissue transglutaminase (TG2) is presented based on aggregation-induced emission (AIE). It has high sensitivity and specificity, minimal background signal and can also effectively distinguish different cell types (drug-resistant cancer cells, cancer cells and normal cells).


Assuntos
Proteínas de Ligação ao GTP/análise , Microscopia Confocal , Transglutaminases/análise , Linhagem Celular , Cisteamina/química , Cisteamina/metabolismo , Resistencia a Medicamentos Antineoplásicos/genética , Corantes Fluorescentes/química , Proteínas de Ligação ao GTP/antagonistas & inibidores , Proteínas de Ligação ao GTP/metabolismo , Humanos , Nanocompostos/química , Peptídeos/química , Peptídeos/metabolismo , Agregados Proteicos , Proteína 2 Glutamina gama-Glutamiltransferase , Transglutaminases/antagonistas & inibidores , Transglutaminases/metabolismo
2.
Biosens Bioelectron ; 126: 249-254, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30445299

RESUMO

Herein, an effective membrane-to-intracellular cholesterol detection strategy was designed based on cascade reactions. A biochip array was firstly fabricated by consecutively immobilizing luminol modified gold nanoparticles (Au@luminol), soybean peroxidase (SBP) and cholesterol oxidase (ChoX) on the cellulose acetate (CA) membrane functionalized home-made micropore array. When cholesterol existed, it was oxidized by ChoX generating H2O2, which further triggered the CL reaction under the SBP catalysis, the CL signals were collected by a charge-coupled device (CCD). The proposed strategy exhibited a wide linear range from 0.12 µM to 1000 µM and relatively low detection limit (LOD) of 0.08 µM. Furthermore,it could be used to in-situ detect membrane cholesterol and intracelluar esterified cholesterol in HepG2 cells. After activated HepG2 cells were added to the modified biochip, membrane cholesterol was detected directly. Intracelluar esterified cholesterol was detected through the introduction of triton X-100 and cholesteryl esterase (ChoE). Additionally, the cholesterol content in cells was changed after stimulated by drugs, such as apolipoprotein A-I (ApoA-I), pitavastatin or probucol. The correlation of the CL signal with the amount of cholesterol confirmed that our strategy was feasible to simultaneously detect membrane and intracellular cholesterol at different cellular states. The proposed strategy exhibited excellent sensitivity, selectivity, stability, and reproducibility in a simple, cheap way, which opened a new door for studying clinic treatment of the cholesterol-related diseases.


Assuntos
Técnicas Biossensoriais , Colesterol Oxidase/química , Colesterol/isolamento & purificação , Nanopartículas Metálicas/química , Catálise , Colesterol/química , Citoplasma/química , Ouro , Humanos , Peróxido de Hidrogênio/química , Oxirredução
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