Electric eel acetylcholinesterase

Manufactured by Merck Group

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Electric eel acetylcholinesterase is an enzyme extracted from the electric eel. It is used in biochemical research to study the function and regulation of acetylcholinesterase, an important enzyme in the nervous system.

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Acetylcholinesterase (113 citations) Electric eel (5 citations) Electric eel acetylcholinesterase (type-VI-S, (4 citations) Acetylcholinesterase from Electrophorus electricus (electric eel) (2 citations) Acetylcholinesterase (AChE) from electric eel (2 citations) Acetylcholinesterase from electric eel (2 citations)

5 protocols using electric eel acetylcholinesterase

Cholinesterase Inhibition and BACE1 Assay

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Electric eel acetylcholinesterase (AChE, EC3.1.1.7), horse serum butyrylcholinesterase (BChE, EC 3.1.1.8), acetylthiocholine iodide (ACh), butyrylthioline chloride (BCh), 5,5′-dithobis [2-nitrobenzoic acid] (DTNB), ethylenediaminetetraacetic acid (EDTA), berberine, and quercetin were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). The BACE1 FREF assay kit (β-secretase) was purchased from Pan Vera Co. (Madison, WI, USA). ONOO⁻ was purchased from Molecular Probes Cayman (Ann Arbor, MI, USA). All other chemicals and solvents used were purchased from E. Merck, Fluka, or Sigma- Aldrich, unless otherwise stated.

Ali M.Y., Seong S.H., Reddy M.R., Seo S.Y., Choi J.S, & Jung H.A. (2017). Kinetics and Molecular Docking Studies of 6-Formyl Umbelliferone Isolated from Angelica decursiva as an Inhibitor of Cholinesterase and BACE1. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(10), 1604.

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Cholinesterase Inhibition Assay

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Electric eel acetylcholinesterase (AChE, EC 3.1.1.7), equine serum butyrylcholinesterase (BChE, EC 3.1.1.8), 5,5’-dithio-bis-2-nitrobenzoic acid (DTNB), and sodium phosphate dibasic (Na₂HPO₄) were purchased from Sigma-Aldrich (St. Louis, MO, USA). S-butyrylthiocholine chloride (BTCC) from Sigma-Aldrich (Buchs, Switzerland), acetylthiocholine iodide (ATCI) from Sigma-Aldrich (Dorset, UK) and sodium phosphate monobasic (NaH₂PO₄) from Sigma-Aldrich (Munich, Germany) were purchased. Tacrine hydrochloride from Cayman Chemical Company was used as the standard drug, and absolute ethanol from Merck Emsure^® Germany, was used as the solvent. The reagents and solvents for the synthesis work were obtained commercially from Sigma-Aldrich Corporation (St. Louis, MO, USA) and used without any further purification.

Kwong H.C., Mah S.H., Chia T.S., Quah C.K., Lim G.K, & Kumar C.S. (2017). Cholinesterase Inhibitory Activities of Adamantyl-Based Derivatives and Their Molecular Docking Studies. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(6), 1005.

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Ellman's Method for Cholinesterase Inhibition

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Ellman’s colorimetric method with minor modification was employed in the determination of AChE and BChE inhibitory activities^{16 (link)}. Electric eel acetylcholinesterase (AChE, EC 3.1.1.7), equine serum butyrylcholinesterase (BChE, EC 3.1.1.8), 5,5′-Dithiobis(2-nitrobenzoic acid) (DTNB) and sodium phosphate dibasic (Na₂HPO₄) were purchased from Sigma-Aldrich (USA). S-butyrylthiocholine chloride (BTCC) from Sigma-Aldrich (Switzerland), acetylthiocholine iodide (ATCI) from Sigma-Aldrich (United Kingdom) and sodium phosphate monobasic (NaH₂PO₄) from Sigma-Aldrich (Germany) were purchased. Tacrine hydrochloride was purchased from Cayman Chemical Company and absolute ethanol from Merck Emsure® Germany was used as the solvent. Ethanol was used to replace the sample as a control in this assay³¹. Tacrine hydrochloride was used as the reference drug in the inhibition assays of AChE and BChE.

Kwong H.C., Chidan Kumar C.S., Mah S.H., Mah Y.L., Chia T.S., Quah C.K., Lim G.K, & Chandraju S. (2019). Crystal Correlation Of Heterocyclic Imidazo[1,2-a]pyridine Analogues and Their Anticholinesterase Potential Evaluation. Scientific Reports, 9, 926.

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Cholinesterase Activity Assay

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Acetylcholine iodide, butyrylcholine
iodide, 5,5-dithio-bis-nitrobenzoic acid (DTNB), electric eel acetylcholinesterase
(type-VI-S), equine butyrylcholinesterase, 2,2-diphenyl-1-picrylhydrazyl
(DPPH), 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid
(ABTS), HPLC-grade methanol, donepezil, and scopolamine were purchased
from Sigma-Aldrich. All chemicals and solvents used were of analytical
grades.

Karim N., Khan I., Khan I., Halim S.A., Khalid A., Abdalla A.N., Rehman N.U., Khan A, & Al-Harrasi A. (2023). Antiamnesic Effects of Novel Phthalimide Derivatives in Scopolamine-Induced Memory Impairment in Mice: A Useful Therapy for Alzheimer’s Disease. ACS Omega, 8(8), 8052-8065.

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Organophosphate Probe Handling and Characterization

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Considering previous observations that fluorophosphonate probes hydrolyze when stored under neat conditions,^{29 (link)} probes were dissolved in anhydrous DMSO shortly after synthesis and characterization and divided into aliquots for longer term storage, avoiding freeze–thaw cycles.
Organophosphates were purchased from the following suppliers and used without further purification: paraoxon (Chem Cruz, sc-208151, Lot D2117) and chlorpyrifos oxon (98.8%, Chem Service, MET-114590, Lot 525430). Stock solutions of OPs were prepared in dry DMSO and stored at −70 °C, avoiding freeze–thaw cycles. 2-Pralidoxime chloride (2-PAM) was purchased from Sigma-Aldrich. Stock solutions of 2-PAM were prepared freshly on the day of the experiment in Milli-Q water and maintained on ice.
Electric eel acetylcholinesterase (Sigma-Aldrich) was prepared at 2 mg/mL concentration in 100 mM PBS, pH 7.4, with 1 mg/mL bovine serum albumin (BSA) and stored at 4 °C for up to 3 months.
Protein concentrations were determined using a bicinchoninic acid (BCA) assay (Thermo Scientific Pierce). Colorimetric assays were performed using a Molecular Devices plate reader.
Synthetic procedures are described in the Supporting Information.

Lin V.S., Volk R.F., DeLeon A.J., Anderson L.N., Purvine S.O., Shukla A.K., Bernstein H.C., Smith J.N, & Wright A.T. (2020). Structure Dependent Determination of Organophosphate Targets in Mammalian Tissues Using Activity-Based Protein Profiling. Chemical research in toxicology, 33(2), 414-425.

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