A practical strategy to design and develop an isoform-specific fluorescent probe for a target enzyme: CYP1A1 as a case study

doi:10.1039/c6sc03970g

	A practical strategy to design and develop an isoform-specific fluorescent probe for a target enzyme: CYP1A1 as a case study
	Ge, Guang-Bo 1,3; Dai, Zi-Ru 1,2; Feng, Lei 3; Jin, Qiang 1,2; Cheng, Hailing 4; Li, Yan 1; Ning, Jing 1; Yu, Yang 1; Cui, Jing-Nan 3; Yang, Ling 1
刊名	CHEMICAL SCIENCE
	2017
卷号	8 期号:4 页码:2795-2803
ISSN号	2041-6520
DOI	10.1039/c6sc03970g
文献子类	Article
英文摘要	The development of isoform-specific probe(s) for a target enzyme with multiple homologs is always challenging. Herein, a practical strategy was used to design and develop an isoform-specific probe for CYP1A1, a key cytochrome P450 isoenzyme involved in xenobiotic metabolism and bioactivation. On the basis of the subtle differences in 3D structure and substrate preference between CYP1A1 and its homolog CYP1A2, we proposed that it was possible to design a CYP1A1-specific probe via local modification of the reaction site on known CYP1A substrates. To validate this hypothesis, 4-hydroxy-1,8-naphthalimide (HN) was selected as the basic fluorophore due to its excellent optical properties, while a series of O-alkylated HN derivatives were prepared to evaluate their specificity towards CYP1A1. Our results revealed that the introduction of a chloroethyl to HN could get the best isoform selectivity towards CYP1A1 over other CYPs including CYP1A2. The newly developed probe NBCeN exhibited excellent specificity, high sensitivity, and a ratiometric fluorescence response following CYP1A1-catalyzed O-dechloroethylation. NBCeN was successfully used to real-time monitor the activity of CYP1A1 in complex biological samples and to rapidly screen CYP1A1 modulators in living systems. NBCeN could also be used for two-photon imaging of intracellular CYP1A1 in living cells and tissues with high ratiometric imaging resolution and deep tissue penetration. All these findings demonstrated that local modification of non-specific substrates was a practical strategy to develop an isoform-specific probe for a target isoenzyme, while NBCeN could serve as a specific imaging tool to explore the biological functions of CYP1A1 in complex biological systems.
WOS关键词	POLYCYCLIC AROMATIC-HYDROCARBONS ; HUMAN CYTOCHROME-P450 1A2 ; HUMAN CARBOXYLESTERASE 1 ; MESSENGER-RNA LEVELS ; XENOBIOTIC METABOLISM ; HUMAN HEPATOCYTES ; HUMAN PROTEOME ; LIVING CELLS ; HUMAN LIVER ; HUMAN LUNG
WOS研究方向	Chemistry
语种	英语
出版者	ROYAL SOC CHEMISTRY
WOS记录号	WOS:000397560500037
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/169382]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Ge, Guang-Bo; Yang, Ling
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China 3.Dalian Univ Technol, State Key Lab Fine Chem, Dalian, Peoples R China 4.Dalian Med Univ, Hosp 2, Inst Canc, Dalian, Peoples R China
推荐引用方式 GB/T 7714	Ge, Guang-Bo,Dai, Zi-Ru,Feng, Lei,et al. A practical strategy to design and develop an isoform-specific fluorescent probe for a target enzyme: CYP1A1 as a case study[J]. CHEMICAL SCIENCE,2017,8(4):2795-2803.
APA	Ge, Guang-Bo.,Dai, Zi-Ru.,Feng, Lei.,Jin, Qiang.,Cheng, Hailing.,...&Yang, Ling.(2017).A practical strategy to design and develop an isoform-specific fluorescent probe for a target enzyme: CYP1A1 as a case study.CHEMICAL SCIENCE,8(4),2795-2803.
MLA	Ge, Guang-Bo,et al."A practical strategy to design and develop an isoform-specific fluorescent probe for a target enzyme: CYP1A1 as a case study".CHEMICAL SCIENCE 8.4(2017):2795-2803.