title: In situ formation of metal coordination polymer: a strategy for fluorescence turn-on assay of acetylcholinesterase activity and inhibitor screening.
abstract: A novel method for the sensing of acetylcholinesterase (AChE) activity and inhibitor screening based on the formation of metal coordination polymer has been developed. Acetylthiocholine (ATCh) was selected as the substrate. In the presence of AChE, ATCh was hydrolyzed to thiocholine and acetate. Thiocholine interacted with Ag(I) to form a metal coordination polymer. A positively charged perylene probe (probe 1) was employed. The fluorescence of probe 1 was very efficiently quenched by a polyanion [PVS, poly(vinyl sulfonate)]. In the presence of acetylcholinesterase, the positively charged metal coordination polymer newly formed in situ would interact with PVS, probe 1 monomer molecules were released, and a turn on fluorescence signal was detected. The assay is highly sensitive, a limit of detection of 0.04 mU/mL AChE was obtained. The assay is also highly selective, a number of potential interference proteins (enzymes) were tested, and none of them show noticeable interference. Sensing of AChE inhibitor was also demonstrated. Our assay is fairly simple and inexpensive. We envision that it could be used for the sensitive detection of other hydrolytic enzyme activities with properly selected substrates and for the screening of potential inhibitor drugs.
authors: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, PR China.
authors: Dongli
authors: D
authors: Liao
authors: Jian
authors: J
authors: Chen
authors: Huipeng
authors: H
authors: Zhou
authors: Yan
authors: Y
authors: Wang
authors: Yongxin
authors: Y
authors: Li
authors: Cong
authors: C
authors: Yu
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism*; D000122:Acetylthiocholine / Q000378:metabolism; D000818:Animals; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004593:Electrophorus; D057075:Enzyme Assays / Q000379:methods*; D009942:Organometallic Compounds / Q000737:chemistry* / Q000494:pharmacology*; D011145:Polyvinyls / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013451:Sulfonic Acids / Q000737:chemistry*
chemical_list: D002800:Cholinesterase Inhibitors; D009942:Organometallic Compounds; D011145:Polyvinyls; D013451:Sulfonic Acids; C008959:lyapolate; D000122:Acetylthiocholine; D000110:Acetylcholinesterase