Acetylcholinesterase Explained

acetylcholinesterase
Ec Number:	3.1.1.7
Cas Number:	9000-81-1
Go Code:	0003990

Acetylcholinesterase (HGNC symbol ACHE; EC 3.1.1.7; systematic name acetylcholine acetylhydrolase), also known as AChE, AChase or acetylhydrolase, is the primary cholinesterase in the body. It is an enzyme that catalyzes the breakdown of acetylcholine and some other choline esters that function as neurotransmitters:

acetylcholine + H₂O = choline + acetate

It is found at mainly neuromuscular junctions and in chemical synapses of the cholinergic type, where its activity serves to terminate synaptic transmission. It belongs to the 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 is a hydrolase that hydrolyzes choline esters. It has a very high catalytic activity—each molecule of AChE degrades about 5,000 molecules of acetylcholine (ACh) per second,^[1] approaching the limit allowed by diffusion of the substrate.^{[2] [3]} The active site of AChE comprises two 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.^{[4] [5]}

The anionic subsite accommodates the positive quaternary 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 aromatic residues that line a gorge leading to the active site.^{[6] [7] [8]} All 14 amino acids in the aromatic gorge are highly conserved across different species.^[9] Among the aromatic amino acids, tryptophan 84 is critical and its substitution with alanine results in a 3000-fold decrease in reactivity.^[10] The gorge is approximately 20 angstroms deep and five angstroms wide.^[11]

The esteratic subsite, where acetylcholine is hydrolyzed to acetate and choline, contains the catalytic triad of three amino acids: serine 203, histidine 447 and glutamate 334. These three amino acids are similar to the triad in other serine proteases except that the glutamate is the third member rather than aspartate. Moreover, the triad is of opposite chirality to that of other proteases.^[12] The hydrolysis reaction of the carboxyl ester leads to the formation of an acyl-enzyme and free choline. Then, the acyl-enzyme undergoes nucleophilic attack by a water molecule, assisted by the histidine 440 group, liberating acetic acid and regenerating the free enzyme.^{[13] [14]}

Species

AChE is found in many biological species, including humans and other mammals, non-vertebrates, and plants.^{[15] [16] [17] [18]}

In humans, AChE is a cholinergic enzyme involved in the hydrolysis of the neurotransmitter acetylcholine (ACh) into its constituents, choline, and acetate.^[15] Overall, in mammals, AChE is primarily involved in the termination of impulse transmission at cholinergic synapses by rapid hydrolysis of the neurotransmitter acetylcholine.^[15] In non-vertebrates, AChE plays a similar role in nerve conduction processes at the neuromuscular junction. It is usually located in the membranes of these animals and controls ionic currents in excitable membranes.^{[17] [18]}

In plants, the biological functions of AChE are less clear, and its existence has been recognized by indirect evidence of its activity. For instance, a study on Solanum lycopersicum (tomato) identified 87 SlAChE genes containing GDSL lipase/acylhydrolase domain. The study also showed up-and down-regulation of SlAChE genes under salinity stress condition.^[15]

Some marine fungi have been found to produce compounds that inhibit AChE. However, the specific role and mechanisms of AChE in fungi are not as well-studied as in mammals.^[18] The presence and role of AChE in bacteria is not well-documented.^[18]

Biological function

During neurotransmission, ACh is released from the presynaptic neuron into the synaptic cleft and binds to ACh receptors on the post-synaptic membrane, relaying the signal from the nerve. AChE is concentrated in the synaptic cleft, where it terminates the signal transmission by hydrolyzing ACh. The liberated choline is taken up again by the pre-synaptic neuron and ACh is synthesized by combining with acetyl-CoA through the action of choline acetyltransferase.^{[19] [20]}

A cholinomimetic drug disrupts this process by acting as a cholinergic neurotransmitter that is impervious to acetylcholinesterase's lysing action.

Disease relevance

See main article: Acetylcholinesterase inhibitor. Drugs or toxins that inhibit AChE lead to persistence of high concentrations of ACh within synapses, leading to increased cholinergic signaling within the central nervous system, autonomic ganglia and neuromuscular junctions.^[21]

Irreversible inhibitors of AChE may lead to muscular paralysis, convulsions, bronchial constriction, and death by asphyxiation. Organophosphates (OP), esters of phosphoric acid, are a class of irreversible AChE inhibitors.^[22] 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 covalently bound. Irreversible AChE inhibitors have been used in insecticides (e.g., malathion) and nerve gases for chemical warfare (e.g., Sarin and VX). Carbamates, esters of N-methyl carbamic acid, are AChE inhibitors that hydrolyze in hours and have been used for medical purposes (e.g., physostigmine for the treatment of 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 donepezil are FDA-approved to improve cognitive function in Alzheimer's disease. Rivastigmine is also used to treat Alzheimer's and Lewy body dementia, and pyridostigmine bromide is used to treat myasthenia gravis.^{[23] [24] [25] [26] [27] [28]}

An endogenous inhibitor of AChE in neurons is 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.^[29]

It has also been shown that the main active ingredient in cannabis, tetrahydrocannabinol, is a competitive inhibitor of acetylcholinesterase.^[30]

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.^{[31] [32] [33]}

Acetylcholinesterase is also found on the red blood cell membranes, where different forms constitute the Yt blood group antigens.^[34] Acetylcholinesterase exists in multiple molecular forms, which possess similar catalytic properties, but differ in their 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. Note higher vertebrates also encode a closely related paralog BCHE (butyrylcholinesterase) with 50% amino acid identity to ACHE.^[35] Diversity in the transcribed products from the sole mammalian gene arises from alternative mRNA splicing and post-translational associations of catalytic and structural subunits. There are three known forms: T (tail), R (read through), and H (hydrophobic).^[36]

AChE_T

The major form of acetylcholinesterase found in brain, muscle, and other tissues, known as is the hydrophilic species, which forms disulfide-linked oligomers with collagenous, or 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, ColQ for the neuromuscular junction and PRiMA for synapses.

AChE_H

The other, alternatively spliced form expressed primarily in the erythroid tissues, differs at the C-terminus, and contains a cleavable hydrophobic peptide with a PI-anchor site. It associates with membranes through the phosphoinositide (PI) moieties added post-translationally.^[37]

AChE_R

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.^[38]

Nomenclature

The nomenclatural variations of ACHE and of cholinesterases generally are discussed at Cholinesterase § Types and nomenclature.

Inhibitors

See main article: Acetylcholinesterase inhibitor. For acetylcholine esterase (AChE), reversible inhibitors are those that do not irreversibly bond to and deactivate AChE.^[39] Drugs that reversibly inhibit acetylcholine esterase are being explored as treatments for Alzheimer's disease and myasthenia gravis, among others. Examples include tacrine and donepezil.^[40]

Exposure to acetylcholinesterase inhibitors is one of several studied explanations for the chronic cognitive symptoms veterans displayed after returning from the Gulf War. Soldiers were dosed with AChEI pyridostigmine bromide (PB) as protection from nerve agent weapons. Studying acetylcholine levels using microdialysis and HPLC-ECD, researchers at the University of South Carolina School of Medicine determined PB, when combined with a stress element can lead to cognitive responses.^[41]

• Phytocannabinoids^[42]
  • Cannabidiol
  • Δ⁸-Tetrahydrocannabinol
  • Cannabigerol
  • Cannabigerolic acid
  • Cannabicitran
  • Cannabidivarin
  • Cannabichromene
  • Cannabinol

See also

• Cholinesterases

Further reading

• Silman I, Futerman AH . Modes of attachment of acetylcholinesterase to the surface membrane . Eur. J. Biochem. . 170 . 1–2 . 11–22 . 1988 . 3319614 . 10.1111/j.1432-1033.1987.tb13662.x . free .
• Sussman JL, Harel M, Frolow F, Oefner C, Goldman A, Toker L, Silman I . Atomic structure of acetylcholinesterase from Torpedo californica: a prototypic acetylcholine-binding protein . Science . 253 . 5022 . 872–9 . 1991 . 1678899 . 10.1126/science.1678899 . 1991Sci...253..872S . 28833513 .
• Soreq H, Seidman S . Acetylcholinesterase--new roles for an old actor . Nature Reviews Neuroscience . 2 . 4 . 294–302 . 2001 . 11283752 . 10.1038/35067589 . 5947744 .
• Shen T, Tai K, Henchman RH, McCammon JA . Molecular dynamics of acetylcholinesterase . Acc. Chem. Res. . 35 . 6 . 332–40 . 2003 . 12069617 . 10.1021/ar010025i .
• Pakaski M, Kasa P . Role of acetylcholinesterase inhibitors in the metabolism of amyloid precursor protein . Current Drug Targets. CNS and Neurological Disorders . 2 . 3 . 163–71 . 2003 . 12769797 . 10.2174/1568007033482869 .
• Meshorer E, Soreq H . Virtues and woes of AChE alternative splicing in stress-related neuropathologies . Trends Neurosci. . 29 . 4 . 216–24 . 2006 . 16516310 . 10.1016/j.tins.2006.02.005 . 18983474 .
• Ehrlich G, Viegas-Pequignot E, Ginzberg D, Sindel L, Soreq H, Zakut H . Mapping the human acetylcholinesterase gene to chromosome 7q22 by fluorescent in situ hybridization coupled with selective PCR amplification from a somatic hybrid cell panel and chromosome-sorted DNA libraries . Genomics . 13 . 4 . 1192–7 . 1992 . 1380483 . 10.1016/0888-7543(92)90037-S .
• Spring FA, Gardner B, Anstee DJ . Evidence that the antigens of the Yt blood group system are located on human erythrocyte acetylcholinesterase . Blood . 80 . 8 . 2136–41 . 1992 . 1391965 . 10.1182/blood.V80.8.2136.2136. free .
• Shafferman A, Kronman C, Flashner Y, Leitner M, Grosfeld H, Ordentlich A, Gozes Y, Cohen S, Ariel N, Barak D . Mutagenesis of human acetylcholinesterase. Identification of residues involved in catalytic activity and in polypeptide folding . J. Biol. Chem. . 267 . 25 . 17640–8 . 1992 . 10.1016/S0021-9258(19)37091-7 . 1517212 . free .
• Getman DK, Eubanks JH, Camp S, Evans GA, Taylor P . The human gene encoding acetylcholinesterase is located on the long arm of chromosome 7 . Am. J. Hum. Genet. . 51 . 1 . 170–7 . 1992 . 1609795 . 1682883 .
• Li Y, Camp S, Rachinsky TL, Getman D, Taylor P . Gene structure of mammalian acetylcholinesterase. Alternative exons dictate tissue-specific expression . J. Biol. Chem. . 266 . 34 . 23083–90 . 1992 . 10.1016/S0021-9258(18)54466-5 . 1744105 . free .
• Velan B, Grosfeld H, Kronman C, Leitner M, Gozes Y, Lazar A, Flashner Y, Marcus D, Cohen S, Shafferman A . The effect of elimination of intersubunit disulfide bonds on the activity, assembly, and secretion of recombinant human acetylcholinesterase. Expression of acetylcholinesterase Cys-580----Ala mutant . J. Biol. Chem. . 266 . 35 . 23977–84 . 1992 . 10.1016/S0021-9258(18)54380-5 . 1748670 . free .
• Soreq H, Ben-Aziz R, Prody CA, Seidman S, Gnatt A, Neville L, Lieman-Hurwitz J, Lev-Lehman E, Ginzberg D, Lipidot-Lifson Y . Molecular cloning and construction of the coding region for human acetylcholinesterase reveals a G + C-rich attenuating structure . Proceedings of the National Academy of Sciences of the United States of America . 87 . 24 . 9688–92 . 1991 . 2263619 . 55238 . 10.1073/pnas.87.24.9688 . 1990PNAS...87.9688S . free .
• Chhajlani V, Derr D, Earles B, Schmell E, August T . Purification and partial amino acid sequence analysis of human erythrocyte acetylcholinesterase . FEBS Lett. . 247 . 2 . 279–82 . 1989 . 2714437 . 10.1016/0014-5793(89)81352-3 . 41843002 . 1989FEBSL.247..279C .
• Lapidot-Lifson Y, Prody CA, Ginzberg D, Meytes D, Zakut H, Soreq H . Coamplification of human acetylcholinesterase and butyrylcholinesterase genes in blood cells: correlation with various leukemias and abnormal megakaryocytopoiesis . Proceedings of the National Academy of Sciences of the United States of America . 86 . 12 . 4715–9 . 1989 . 2734315 . 287342 . 10.1073/pnas.86.12.4715 . 1989PNAS...86.4715L . free .
• Bazelyansky M, Robey E, Kirsch JF . Fractional diffusion-limited component of reactions catalyzed by acetylcholinesterase . Biochemistry . 25 . 1 . 125–30 . 1986 . 3954986 . 10.1021/bi00349a019 .
• Gaston SM, Marchase RB, Jakoi ER . Brain ligatin: a membrane lectin that binds acetylcholinesterase . J. Cell. Biochem. . 18 . 4 . 447–59 . 1982 . 7085778 . 10.1002/jcb.1982.240180406 . 22975039 .
• Ordentlich A, Barak D, Kronman C, Ariel N, Segall Y, Velan B, Shafferman A . Contribution of aromatic moieties of tyrosine 133 and of the anionic subsite tryptophan 86 to catalytic efficiency and allosteric modulation of acetylcholinesterase . J. Biol. Chem. . 270 . 5 . 2082–91 . 1995 . 7836436 . 10.1074/jbc.270.5.2082 . free .
• Maruyama K, Sugano S . Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides . Gene . 138 . 1–2 . 171–4 . 1994 . 8125298 . 10.1016/0378-1119(94)90802-8 .
• Ben Aziz-Aloya R, Sternfeld M, Soreq H . Promoter elements and alternative splicing in the human ACHE gene . Prog. Brain Res. . 98 . 147–53 . 1994 . 8248502 . 10.1016/s0079-6123(08)62392-4 .
• Massoulié J, Pezzementi L, Bon S, Krejci E, Vallette FM . Molecular and Cellular Biology of Cholinesterases . Prog. Brain Res. . 41 . 1 . 31–91 . 1993 . 8321908 . 10.1016/0301-0082(93)90040-Y . 21601586 .

External links

• ATSDR Case Studies in Environmental Medicine: Cholinesterase Inhibitors, Including Insecticides and Chemical Warfare Nerve Agents U.S. Department of Health and Human Services
  • • • • Acetylcholinesterase: A gorge-ous enzyme—PDBe
• Acetylcholinesterase—RCSB PDB

Notes and References

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