title: A direct assay of butyrylcholinesterase activity using a fluorescent substrate.
abstract: In this study, we report a direct fluorometric assay for butyrylcholinesterase (BChE) activity and screening of its inhibitor, using a fluorescent substrate. 2-(2-(5,6-Dimethoxy-1,3-dioxoisoindolin-2-yl)acetoxy)-N,N,N-trimethylethan-1-ammonium iodide (1) was hydrolyzed by BChE, and its fluorescence was quenched by an intramolecular photoinduced electron transfer process. The resulting change in fluorescence provided a facile method for real-time BChE activity testing. Remarkably, 1 was selectively hydrolyzed by BChE, even in the presence of excess acetylcholinesterase, thereby facilitating the specific monitoring of BChE activity. This assay method is also useful for screening potential BChE inhibitors. Given its simplicity, selectivity, and higher assay speed, this method may be extended to high-throughput screening of BChE inhibitors and relevant drug discovery.
authors.affiliation: Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Republic of Korea. happyhan@gist.ac.kr.
authors.forename: Seungyoon
authors.initials: S
authors.lastname: Kang
authors.affiliation: Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Republic of Korea. happyhan@gist.ac.kr.
authors.forename: Suji
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Republic of Korea. happyhan@gist.ac.kr.
authors.forename: Woojin
authors.initials: W
authors.lastname: Yang
authors.affiliation: Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Republic of Korea. happyhan@gist.ac.kr.
authors.forename: Jiwon
authors.initials: J
authors.lastname: Seo
authors.affiliation: Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Republic of Korea. happyhan@gist.ac.kr.
authors.forename: Min Su
authors.initials: MS
authors.lastname: Han
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D002091:Butyrylcholinesterase / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004593:Electrophorus; D057075:Enzyme Assays / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism*; D005470:Fluorometry / Q000379:methods; D006736:Horses; D006868:Hydrolysis / Q000187:drug effects; D013552:Swine
chemical_list: D002800:Cholinesterase Inhibitors; D005456:Fluorescent Dyes; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase