Acetylcholinesterase Human Enzymatic Inhibition Reversibility LeadHunter Assay - FR

Background Information:

Acetylcholinesterase (ACES, AchE, EC 3.1.1.7)) terminates the action of acetylcholine (ACh) at the junctions of various cholinergic nerve endings with their effector organs or postsynaptic sites. Compounds that inhibit AChE are called anticholinesterase agents and are used to ameliorate the symptoms of myesthenia gravis, lower intraocular pressure in glaucoma and for possible treatment of dementia through increase of ACh concentration in the brain. Anticholinesterases may also cause salivation, meiosis, increase in gastrointestinal motility, skeletal muscle paralysis, respiratory arrest and cardiovascular collapse. These agents are also used as insecticides and pesticides for agricultural purposes and vector control.
AChE enzymatic activity is measured by jump dilution using the standard colorimetric assay (Eurofins Cerep ref. 363). The project is done in two step: (1) the determination of IC50 in our standard assay . (2) Then the reversibility assessment is performed. It consists in a rapid and large dilution of the enzyme-inhibitor complex . The enzyme 100-fold-concentrated is pre-incubated (30 minutes) with the test compounds (10 fold IC50), then the mixture is diluted 100-fold into the reaction buffer containing the enzyme substrates to initiate the reaction. The kinetic of the reaction is followed during 12 time points and compared to the control without inhibitor. Diisopropylfluorophosphate is used as irreversible inhibitor reference and Galanthamine is used as a reversible inhibitor reference.

Family:

Neurotransmitter Metabolism

Sub Family:

Acetylcholine Turnover

Protein Name:

Acetylcholinesterase

Uniprot Number:

P22303

Protein Aliases:

Yt blood group

Gene Name:

ACHE

Gene ID:

43

Organism:

Human