title: A highly selective ratiometric fluorescent probe for in vitro monitoring and cellular imaging of human carboxylesterase 1.
abstract: A new ratiometric fluorescent probe derived from 2-(2-hydroxy-3-methoxyphenyl) benzothiazole (HMBT) has been developed for selective monitoring of human carboxylesterase 1 (hCE1). The probe is designed by introducing benzoyl moiety to HMBT. The prepared latent spectroscopic probe 1 displays satisfying stability under physiological pH conditions with very low background signal. Both the reaction phynotyping and chemical inhibition assays demonstrated that hCE1 mediated the specific cleavage of the carboxylic ester bond of probe 1 in human biological samples. The release of HMBT leads to a remarkable red-shifted emission in fluorescence spectrum (120 nm large emission shift). Furthermore, human cell-based assays show that probe 1 is cell membrane permeable, and it can be used for bioassay and cellular imaging of hCE1 activity in HepG2 cells. These findings lead to the development of a simple and sensitive fluorescent method for measurement of hCE1 activity in vitro or in living cells, in the presence of additional enzymes or endogenous compounds.
authors.affiliation: Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian 116023, China.
authors.forename: Zhao-Ming
authors.initials: ZM
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Feng
authors.affiliation: Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian 116023, China.
authors.forename: Guang-Bo
authors.initials: GB
authors.lastname: Ge
authors.affiliation: Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian 116023, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Lv
authors.affiliation: Dalian Medical University, Dalian 116044, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Hou
authors.affiliation: Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian 116023, China.
authors.forename: Yun-Feng
authors.initials: YF
authors.lastname: Cao
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China. Electronic address: jncui@dlut.edu.cn.
authors.forename: Jing-Nan
authors.initials: JN
authors.lastname: Cui
authors.affiliation: Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhong-shan Road, Dalian 116023, China. Electronic address: yling@dicp.ac.cn.
authors.forename: Ling
authors.initials: L
authors.lastname: Yang
mesh_terms: D052160:Benzothiazoles / Q000032:analysis*; D015374:Biosensing Techniques / Q000379:methods; D002265:Carboxylic Ester Hydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D057075:Enzyme Assays / Q000379:methods; D005456:Fluorescent Dyes / Q000032:analysis*; D056945:Hep G2 Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D008862:Microsomes, Liver / Q000201:enzymology; D061848:Optical Imaging / Q000379:methods*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D052160:Benzothiazoles; D005456:Fluorescent Dyes; D002265:Carboxylic Ester Hydrolases; C474609:CES1 protein, human; C005465:benzothiazole
keywords: Cell imaging; Human carboxylesterase 1; Large emission shift; Ratiometric fluorescent probe