Neurotransmitters inactivation mechanisms

Kennedy, C, Todorov, LD, Mihaylova-Todorova, S and Sneddon, P (1997) Release of soluble nucleotidases a novel mechanism for neurotransmitter inactivation. Trends Pharmacol. Sci. 18 263-266. 

Anandamide is inactivated in two steps, first by transport inside the cell and subsequently by intracellular enzymatic hydrolysis. The transport of anandamide inside the cell is a carrier-mediated activity, having been shown to be a saturable, time- and temperature-dependent process that involves some protein with high affinity and specificity for anandamide (Beltramo, 1997). This transport process, unlike that of classical neurotransmitters, is Na+-independent and driven only by the concentration gradient of anandamide (Piomelli, 1998). Although the anandamide transporter protein has not been cloned yet, its well characterized activity is known to be inhibited by specific transporter inhibitors. Reuptake of 2-AG is probably mediated by the same facilitating mechanism (Di Marzo, 1999a,b Piomelli, 1999). 

Almost invariably, a neuron is genetically programmed to synthesize and release only a single type of neurotransmitter. Therefore, a given synapse is either always excitatory or always inhibitory. Once a neurotransmitter has bound to its receptor on the postsynaptic neuron and has caused its effect, it is important to inactivate or remove it from the synapse in order to prevent its continuing activity indefinitely. Several mechanisms to carry this out have been identified ... 

For any substance to serve effectively as a neurotransmitter, it must be rapidly removed or inactivated from the synapse or, in this case, the neuroeffector junction. This is necessary in order to allow new signals to get through and influence effector tissue function. Neurotransmitter activity may be terminated by three mechanisms ... 

The primary mechanism used by cholinergic synapses is enzymatic degradation. Acetylcholinesterase hydrolyzes acetylcholine to its components choline and acetate it is one of the fastest acting enzymes in the body and acetylcholine removal occurs in less than 1 msec. The most important mechanism for removal of norepinephrine from the neuroeffector junction is the reuptake of this neurotransmitter into the sympathetic neuron that released it. Norepinephrine may then be metabolized intraneuronally by monoamine oxidase (MAO). The circulating catecholamines — epinephrine and norepinephrine — are inactivated by catechol-O-methyltransferase (COMT) in the liver. 

Because duration of activity of the catecholamines is significantly longer than that of neuronally released norepinephrine, the effects on tissues are more prolonged. This difference has to do with the mechanism of inactivation of these substances. Norepinephrine is immediately removed from the neuroeffector synapse by way of reuptake into the postganglionic neuron. This rapid removal limits duration of the effect of this neurotransmitter. In... 

In contrast, pertussis toxin catalyzes the ADP-ribosyl-ation of a specific cysteine residue in Gai) G(m and Gal [1]. Only a subunits bound to their Py subunits can undergo this modification. Pertussis-toxin-mediated ADP-ribosylation inactivates these a subunits such that they cannot exchange GTP for GDP in response to receptor activation (Fig. 19-1B). By this mechanism, pertussis toxin blocks the ability of neurotransmitters to inhibit adenylyl cyclase or to influence the gating of K+ and Ca2+ channels in target neurons. However, since G is not a substrate for pertussis toxin, the toxin may not be able to block neurotransmitter-mediated inhibition of adenylyl cyclase in all cases. The Gq and Gn 16 types of G protein a subunit are not known to undergo ADP-ribosylation. 

A synaptic neurotransmitter may become inactivated by several mechanisms. It may passively diffuse out of the synapse it may undergo enzymatic breakdown it may be taken up into a cell through a selective reuptake pump and, intracellularly, it may be taken back into a vesicle for future release, or it may be converted enzymatically. 

Activation of K+ channels, inactivation of Ca2+ channels and direct inhibition of neurotransmitter release are powerful mechanisms by which opioids inhibit the neuronal transmission of the pain signal. 

These reactions, which have provided a means of inhibiting the flavin-linked monoamine oxidases, enable us to end on a clinical note. The monoamine oxidases are responsible for the deamination of monoamines such as adrenaline, noradrenaline, dopamine, and serotonin, which act as neurotransmitters. Imbalances in the levels of monoamines cause various psychiatric and neurological disorders Parkinson s disease is associated with lowered levels of dopamine, and low levels of other monoamines are associated with depression. Inhibitors of monoamine oxidases may consequently be used to treat Parkinson s disease and depression. The flavin moiety is covalently bound to the enzyme by the thiol group of a cysteine residue (equation 9.17). The acetylenic suicide inhibitor N,N-dimethyl-propargylamine inactivates monoamine oxidases by alkylating the flavin on N-5.25 A likely mechanism for the reaction is the Michael addition of the N-5 of the reduced flavin to the acetylenic carbon 2... 

Neurotransmitters are removed by translocation into vesicles or destroyed in enzyme-catalysed reactions. Acetylcholine must be removed from the synaptic cleft to permit repolarization and relaxation. A high affinity acetylcholinesterase (AChE) (the true or specific AChE) catalyses the hydrolysis of acetylcholine to acetate and choline. A plasma AChE (pseudo-AChE or non-specific AChE) also hydrolyses acetylcholine. A variety of plant-derived substances inhibit AChE and there is considerable interest in AChE inhibitors as potential therapies for cognition enhancement and for Alzheimer s disease. Organophosphorous compounds alkylate an active site serine on AChE and the AChE inhibition by this mechanism is the basis for the use of such compounds as insecticides (and unfortunately also as chemical warfare agents). Other synthetics with insecticidal and medical applications carbamoylate and thus inactivate AChE (Table 6.4). 

Discovery. The majority of both old and new antidepressants act by virtue of their ability to inhibit monoamine transporter mechanisms in the brain. The concept that neurotransmitters are inactivated by uptake of the released chemical into the nerve terminal from which it had been released or into adjacent cells is less than 40 years old. Before this it was generally assumed that the inactivation of norepinephrine and the other monoamine neurotransmitters after their release from nerves was likely to involve rapid enzymatic breakdown, akin to that seen with acetylcholinesterase. The degradation of monoamines by the enzyme monoamine oxidase vas known early on, and in the 1950s a second enzyme catechol-O-methyl transferase (COMT) vas discovered and was thought to play a key role in inactivating norepinephrine and other catecholamines. 

Unfortunately, this is not possible for a number of reasons. Many are not chemically stable enough to survive the acid of the stomach and would have to be injected. Even if they were injected, there is little chance that they would survive to reach their target receptors. As mentioned already, the body has efficient mechanisms by which it inactivates its neurotransmitters as soon as they have passed on their message. Therefore, on injection, they would be swiftly inactivated by enzymes or by cell uptake. 

Mechanisms of Inactivation. It was noted previously that one of the criteria for the identification of a neurotransmitter is the presence of a local means by which a transmitter can be inactivated. If this process were not present, it stands to reason that a transmitter would be exposed to a postsynaptic recognition site for a prolonged period of time, an event that... 

Specific decarboxylases are known for a majority of the amino acids, and several are prime targets for inactivation by virtue of their substantial medicinal importance. These include aromatic-amino-acid decarboxylase, which is responsible for the production of dopamine (DOPA) orithine decarboxylase, which supplies the p amine putrescine and glutamate decarboxylase, which converts glutamate to the inhibitory neurotransmitter y-aminobutyric acid (GABA). The accepted mechanism of these enzymes involves decarboxylation of the amino acid to yield a resonance-stabilized carbanion at the a-carbon of the substrate. The intermediate is then protonated with retention of configuration to yield product (Walsh, 1979, p. 800). 

NEUROTRANSMITTERS Transmitters may produce minimal effects on bioelectric properties, yet activate or inactivate biochemical mechanisms necessary for responses to other circuits. Alternatively, the action of a transmitter may vary with the context of ongoing synaptic events— enhancing excitation or inhibition, rather than operating to impose direct excitation or inhibition. Each chemical substance that fits within the broad definition of a transmitter may therefore require operational definition within the spatial and temporal domains of a specific ceU-ceU circuit. Those same properties may not necessarily be generalized to other cells contacted by the same presynaptic neurons differences in operation may relate to differences in postsynaptic receptors and the mechanisms by which an activated receptor produces its effects. 

The primary mechanism of action of OP pesticides is inhibition of AChE. AChE is an enzyme found in the central nervous system (CNS) and the peripheral nervous system, and its normal physiologic action is to metabolize acetylcholine (ACh), a neurotransmitter. OPs inactivate AChE by phosphorylating the. serine hydroxyl group located at the active site of AChE. The phosphorylation occurs by loss of an OP leaving group and establishment of a covalent bond with AChE. 

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