Acetylcholinesterase: A Representative Inactivation Enzyme

This is the side view of the human brain, facing left. The lights are showing you some of the places where neurons use acetylcholine as their neurotransmitter. We now zoom in on one of these neurons which uses acetylcholine. The cell body, located in the septal nucleus of the hypothalamus, sends its axon to the hippocampus. At the end of the axon is the terminal bouton. After the arrival of an action potential, vesicles containing acetylcholine fuse with the presynaptic membrane and release acetylcholine into the synaptic cleft.

Acetylcholine (ACh), the first neurotransmitter ever to be identified, is a smallmolecule excitatory neurotransmitter with a wide variety of known functions. Within the central nervous system, cholinergic cells (neurons that use ACh as a neurotransmitter) are found in several different locations of the brain, including the striatal complex, the basal forebrain, the diencephalon, pontomesencephalic cell groups, and the medulla. Depending on which area of the brain it is found in, ACh may be involved in any one of several different functions. Some of these functions include the conduction of pain, the regulation of neuroendocrine function, the regulation of REM sleep cycles, and the process of learning and memory formation. In the sympathetic and parasympathetic nervous systems and at all neuromuscular junctions, ACh is used to signal muscle movement. Whether it is within the brain, or at the neuromuscular junction, when ACh is released into the synaptic cleft, it transiently binds with a ligand-gated receptor molecule on the post-synaptic neuron or muscle cell. This binding causes the receptor molecule to open up, allowing for an exchange of ions which subsequently results in a depolarization of the post-synaptic neuron. If this depolarization is strong enough, an action potential is stimulated. After depolarization, the neurotransmitter must be swiftly removed from the receptor molecule and inactivated in order to prevent constant stimulation of the post-synaptic membrane and an excessive firing of action potentials. ACh is one of several neurotransmitters whose action at the synaptic cleft is terminated by the hydrolyizing action of an enzyme. Once ACh has been recognized and the postsynaptic membrane has been depolarized, ACh quickly dissociates from the receptor and binds to acetylcholinesterase (AChE), an enzyme named for its ability to break the ester bond that holds acetate and choline together. Removal of ACh from the recognition site causes the membrane-spanning pore of the receptor to close. The electric potential of the membrane then normalizes and the action potential ceases to fire. The breakdown of ACh by AChE inactivates the neurotransmitter and prevents it from reassociating with the receptor molecule.
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IVA2. Location, Structure and Function of Acetylcholinesterase Presynaptic vesicles release acetylcholine into the synaptic cleft where it binds to its receptor. Right next to the receptor is acetylcholinesterase, the enzyme which breaks up acetylcholine into acetate and choline. AChE is a glycoprotein that exists is several forms. For images of acetylcholinesterase's structure displayed in ribbon form, go tohttp://jsdnt.claremont.edu/biochem98/acetylcholinesterase/AChEcontents.html. As a result of the variety in chemical structure, some forms of AChE are hydrophobic, while others are hydrophilic. The hydrophilic species generally work within the cell to break down excess concentrations of intracellular ACh. The lipidlinked (hydophobic) varieties, however, are the primary agents of ACh inactivation, working at the synaptic cleft or the neuromuscular junction to break ACh into its component parts: acetate and choline. Lipid-linked species of AChE are embedded within the post-synaptic membrane and placed strategically close to the post-synaptic receptor molecules in order to ensure quick inactivation of ACh. Though the individual chemical qualities and anatomical arrangements vary widely between the various forms of AChE, the mechanism of catalysis for all the species remains strikingly similar. In all cases, the active site of AChE is made up of two subsites, both of which are critical to the breakdown of ACh. The anionic site serves to bind a molecule of ACh to the enzyme. When first discovered, this site was thought to be made up of one or more negatively charged groups which electrostatically interacted with the positively charged quaternary nitrogen of the ACh molecule. More recently, however, it is thought that hydrophobic interactions are equally, if not more important in binding this region of the substrate to the enzyme. Once the ACh is bound, the hydrolytic reaction occurs at a second region of the active site called the esteratic subsite. Here, the ester bond of ACh is broken, releasing acetate and choline. Choline is then either temporarily trapped within the junctional folds of the muscular endplate or immediately taken up again by the high affinity choline uptake (HACU) system on the presynaptic membrane. Acetate, however, becomes covalently bonded to serine residues within the esteratic subsite, forming a temporary acetylated form of AChE. A molecule of water then reacts with this intermediate, liberating the acetate group, which diffuses into the surrounding medium. What remains is an active form of the enzyme, ready to react with a newly released molecule of ACh.