Acetylcholinesterase

Acetylcholinesterase, also known as AChE or acetylhydrolase, is a is_associated_with::hydrolase that is_associated_with::hydrolyzes the is_associated_with::neurotransmitter is_associated_with::acetylcholine. AChE is found at mainly is_associated_with::neuromuscular junctions and is_associated_with::cholinergic is_associated_with::brain synapses, where its activity serves to terminate is_associated_with::synaptic transmission. It belongs to is_associated_with::carboxylesterase family of enzymes. It is the primary target of inhibition by organophosphorus compounds such as nerve agents and pesticides.

Enzyme structure and mechanism


AChE has a very high is_associated_with::catalytic activity - each molecule of AChE degrades about 25000 molecules of is_associated_with::acetylcholine (ACh) per second, approaching the limit allowed by is_associated_with::diffusion of the substrate. The is_associated_with::active site of AChE comprises 2 subsites -  the anionic site and the esteratic subsite. The structure and mechanism of action of AChE have been elucidated from the crystal structure of the enzyme.

The anionic subsite accommodates the positive quaternary is_associated_with::amine of acetylcholine as well as other cationic substrates and inhibitors. The cationic substrates are not bound by a negatively charged amino acid in the anionic site, but by interaction of 14 is_associated_with::aromatic residues that line the gorge leading to the active site. All 14 amino acids in the aromatic gorge are highly conserved across different species. Among the aromatic amino acids, is_associated_with::tryptophan 84 is critical and its substitution with is_associated_with::alanine results in a 3000-fold decrease in reactivity. The gorge penetrates half way through the enzyme and is approximately 20 is_associated_with::angstroms long. The active site is located 4 angstroms from the bottom of the molecule.

The esteratic subsite, where acetylcholine is hydrolyzed to acetate and choline, contains the is_associated_with::catalytic triad of three amino acids: is_associated_with::serine 200, is_associated_with::histidine 440 and is_associated_with::glutamate 327. These three amino acids are similar to the triad in other is_associated_with::serine proteases except that the glutamate is the third member rather than is_associated_with::aspartate. Moreover, the triad is of opposite chirality to that of other proteases. The hydrolysis reaction of the carboxyl ester leads to the formation of an acyl-enzyme and free is_associated_with::choline. Then, the acyl-enzyme undergoes is_associated_with::nucleophilic attack by a water molecule, assisted by the histidine 440 group, liberating is_associated_with::acetic acid and regenerating the free enzyme.

Biological function
During is_associated_with::neurotransmission, ACh is released from the nerve into the synaptic cleft and binds to ACh receptors on the post-synaptic membrane, relaying the signal from the nerve. AChE, also located on the post-synaptic membrane, terminates the signal transmission by hydrolyzing ACh. The liberated choline is taken up again by the pre-synaptic nerve and ACh is synthetized by combining with is_associated_with::acetyl-CoA through the action of is_associated_with::choline acetyltransferase.

Disease relevance
For a cholinergic neuron to receive another impulse, ACh must be released from the ACh receptor. This occurs only when the concentration of ACh in the synaptic cleft is very low. Inhibition of AChE leads to accumulation of ACh in the synaptic cleft and results in impeded neurotransmission.



Irreversible inhibitors of AChE may lead to muscular is_associated_with::paralysis, convulsions, is_associated_with::bronchial constriction, and death by is_associated_with::asphyxiation. is_associated_with::Organophosphates (OP), esters of phosphoric acid, are a class of irreversible AChE inhibitors. Cleavage of OP by AChE leaves a phosphoryl group in the esteratic site, which is slow to be hydrolyzed (on the order of days) and can become is_associated_with::covalently bound. Irreversible AChE inhibitors have been used in is_associated_with::insecticides (e.g., is_associated_with::malathion) and nerve gases for chemical warfare (e.g., is_associated_with::Sarin and is_associated_with::Soman). is_associated_with::Carbamates, esters of N-methyl carbamic acid, are AChE inhibitors that hydrolyze in hours and have been used for medical purposes (e.g., is_associated_with::physostigmine for the treatment of is_associated_with::glaucoma). Reversible inhibitors occupy the esteratic site for short periods of time (seconds to minutes) and are used to treat of a range of central nervous system diseases. Tetrahydroaminoacridine (THA) and is_associated_with::donepezil are FDA-approved to improve cognitive function in is_associated_with::Alzheimer's disease. is_associated_with::Rivastigmine is also used to treat Alzheimer's and is_associated_with::Lewy body dementia, and is_associated_with::pyridostigmine bromide is used to treat is_associated_with::myasthenia gravis. Alzheimer disease drugs is_associated_with::donepezil, is_associated_with::galantamine, and is_associated_with::rivastigmine are inhibitors of acetylcholinesterase as well. An endogenous inhibitor of AChE in neurons is is_associated_with::Mir-132 microRNA, which may limit inflammation in the brain by silencing the expression of this protein and allowing ACh to act in an anti-inflammatory capacity.

It has also been shown that the main active ingredient in cannabis, is_associated_with::tetrahydrocannabinol, is a competitive inhibitor of acetylcholinesterase.

Distribution
AChE is found in many types of conducting tissue: nerve and muscle, central and peripheral tissues, motor and sensory fibers, and cholinergic and noncholinergic fibers. The activity of AChE is higher in motor neurons than in sensory neurons.

Acetylcholinesterase is also found on the is_associated_with::red blood cell membranes, where different forms constitute the is_associated_with::Yt blood group is_associated_with::antigens. Acetylcholinesterase exists in multiple molecular forms, which possess similar catalytic properties, but differ in their is_associated_with::oligomeric assembly and mode of attachment to the cell surface.

AChE gene
In mammals, acetylcholinesterase is encoded by a single AChE gene while some invertebrates have multiple acetylcholinesterase genes. Diversity in the transcribed products from the sole mammalian gene arises from alternative mRNA splicing and is_associated_with::post-translational associations of catalytic and structural subunits. There are three known forms: T (tail), R (read through), and H(hydrophobic).

AChET
The major form of acetylcholinesterase found in brain, muscle, and other tissues, known as is the hydrophilic species, which forms disulfide-linked oligomers with is_associated_with::collagenous, or is_associated_with::lipid-containing structural subunits. In the neuromuscular junctions AChE expresses in asymmetric form which associates with ColQ or subunit. In the central nervous system it is associated with PRiMA which stands for Proline Rich Membrane anchor to form symmetric form. In either case, the ColQ or PRiMA anchor serves to maintain the enzyme in the intercellular junction, is_associated_with::ColQ for the neuromuscular junction and PRiMA for synapses.

AChEH
The other, alternatively spliced form expressed primarily in the erythroid tissues, differs at the is_associated_with::C-terminus, and contains a cleavable is_associated_with::hydrophobic is_associated_with::peptide with a PI-anchor site. It associates with membranes through the is_associated_with::phosphoinositide (PI) moieties added post-translationally.

AChER
The third type has, so far, only been found in Torpedo sp. and mice although it is hypothesized in other species. It is thought to be involved in the stress response and, possibly, inflammation.