Comparison of the structural properties of the active site cavities of human and rat monoamine oxidase A and B in their soluble and membrane-bound forms

Structural properties of the active site cavities in human and rat monoamine oxidases (MAOA and MAOB) have been studied in their detergent-purified and outer mitochondrial membrane (OMM) bound forms using a spin-labeled irreversible inhibitor (ParSL) as an active specific spin probe. ParSL has been found to be 5-10-fold more specific for human MAOB (hMAOB) with a Ki of ca. 20 muM, compared to Ki's in the range of 100-200 muM observed for other human and rat MAOs. Solvent accessibilities of the active-site-bound spin probes have been determined by studying the power saturation properties of the spin probe EPR signals in the presence and absence of a polar paramagnetic reagent NiEDDA and by measuring the extent of spin probe reductions on treatment with excess ascorbic acid. Results presented here show that the spin probe bound to the hMAOA active site is ca. 7-8-fold more accessible than in hMAOB. In contrast, the spin probes covalently attached to the two rat enzyme active sites show comparable accessibilities to each other. On comparison of human versus rat enzymes, the active-site-bound spin probes in the two rat MAOs show ca. 40% less accessibilities compared to the same in hMAOA but ca. 4-5-fold higher accessibilities than in hMAOB active site. The present data thus suggests that the structural properties of the active site cavities in rat MAOs are significantly different compared to those in the two human enzymes, which correlates with the differences reported earlier in the inhibitor specificities between human and rat MAOs.     

The effect of electromagnetic radiation on the monoamine oxidase A activity in the rat brain

The effect of the ultralow power pulse-modulated electromagnetic radiation (EMR, power density 10 microW/cm2; carrying frequency 915 MHz; modulating pulses with frequency 2, 4, 6, 8, 12, 16 and 20 Hz) on activity of monoamine oxidase (MAO-A), enzyme involved in the oxidative deamination of monoamines, was investigated. It was established that the increase of activity MAO in hypothalamus reached the maximal meaning at modulation frequency of 6 Hz that corresponded 160% (p < 0.01) of the control level; and at modulation frequency of 20 Hz the decrease of enzyme activity up to 74% (p < 0.01) was found. Mainly the action of ultralow power pulse-modulated EMR on activity of MAO in hippocamp was activating; and the maximal increase of enzyme activity up to 174% (p < 0.01) was registered at modulation frequency of 4 Hz.     

The oxidation of adrenaline and noradrenaline by the two forms of monoamine oxidase from human and rat brain

The selective monoamine oxidase inhibitors clorgyline and (−)-deprenyl were used to study the distribution of monoamine oxidase-A and -B (MAO-A, MAO-B) activities towards (−)-noradrenaline and (+),(−)-adrenaline in homogenates from seven different regions of human brain. The activities towards 5-hydroxytryptamine and 2-phenethylamine, which are essentially specific substrates for the A- and B-forms, respectively, under the conditions used in this work, were also determined. Noradreanline and adrenaline were substrates for both forms of the enzyme in all regions studied. The total MAO activity was found to be highest in the hypothalamus and lowest in the cerebellar cortex. Use of the selective MAO inhibitors clorgyline and (−)-deprenyl also showed adrenaline and noradrenaline to be substrates for both forms of the enzyme in rat brain. In human cerebral cortex and rat brain the two forms were found to have similar K_m-values and maximum velocities towards adrenaline. These values for the two forms were also found to be similar in human cerebral cortex when noradrenaline was used as the substrate. In contrast MAO-A showed a significantly lower K_m and a higher maximum velocity towards noradrenaline in rat brain. These results suggest that the rat may not provide a close model of the human for studies on the effects of MAO inhibitors on brain noradrenaline metabolism.     

Differential inhibition of soluble and membrane-bound acetylcholinesterase forms from mouse brain by choline esters with an acyl moiety of an intermediate size

Differential inhibitions of soluble and membrane-bound acetylcholinesterase forms purified from mouse brain were examined by the comparison of kinetic constants such as a K _m value, a K_ss value (substrate inhibition constant), and IC₅₀ values of active site-selective ligands including choline esters. Membrane-bound acetylcholinesterase form (solubilized only in the presence of detergent) showed lower K_m and K_ss values than soluble acetylcholinesterase form (easily solubilized without detergent). Edrophonium expressed a slightly but significantly (p<0.01) higher inhibition of detergent-soluble acetylcholinesterase form than aqueous-soluble acetylcholinesterase form, while physostigmine inhibited both forms with a similar potency. A remarkable difference in inhibition was observed using choline esters; although choline esters with acyl chain of a short size (acetyl-to butyrylcholine) or a long size (heptanoyl- to decanoylcholine) showed a similar inhibitory potency for two forms of acetylcholinesterase, pentanoylcholine and hexanoylcholine inhibited more strongly aqueous-soluble acetylcholinesterase than detergent-soluble acetylcholinesterase. Thus, it is suggested that the two forms of AChE may be distinguished kinetically by pentanoyl- or hexanoylcholine.This work was supported in part by Agency for Defense Development.     

In vitro andin vivo effect of organophosphate pesticides on monoamine oxidase activity in rat brain

The effects of some organophosphate pesticides, e.g. lebaycid, metacid and metasystox on the monoamine oxidase (MAO) activity in rat brain mitochondria have been studied. These pesticides cause significant inhibition of MAO activityin vitro but have negligible effects on its activityin vivo.     

Effect of benzamide derivatives on rat brain monoamine oxidase activity in vitro

The ability of moclobamide and other benzamide derivatives to inhibit the activity of monoamine oxidase in the rat brain was studied. Distinct effects of these compounds on the deamination of serotonin and norepinephrine (MAO-A substrates); 2-phenylethylamine (selective MAO-B substrate); tyramine and dopamine (MAO-A and MAO-B substrates) are shown. It was demonstrated that among all the compounds studied moclobamide appeared to be the most active and selective inhibitor of MAO-A: at a concentration of 100 microM it caused a 100% inhibition of serotonin and norepinephrine deamination, which might be explained by the presence of C1 atom in the para-position of benzene ring in moclobamide molecule. Other benzamide derivatives were less active in inhibiting MAO-A and had but a negligible effect on dopamine- and 2-phenylethylamine deamination.     

Influence of age on brain vascular and cardiovascular monoamine oxidase activity in the rat

Monoamine oxidase (MAO) activity in brain microvessels and cardiovascular tissues was examined in rats of different age. MAO activity continued to increase with age in the heart, but in contrast, reached maximum activity in three weeks in the aorta, mesenteric artery and mesenteric vein. Between 7 and 60 weeks, there was a small decline in the MAO activity in the testicular artery. The highest MAO activity was found in the cerebral microvessels and increased with age. The half-life of MAO was estimated in the heart and peripheral blood vessels in young and old animals. The half-life of cardiac MAO was increased with age whereas that of the mesenteric vein, mesenteric artery and aorta remained constant between 7 and 112 weeks. Thus an explanation for this increased cardiac MAO activity in old rats was a reduced rate of degredation of this enzyme. The high activity of the enzyme in the brain microvessels suggests that it may participate in regulating the influx and efflux of monoamines in the central nervous system.     

Effect of heat and cold exposure on the rat brain monoamine oxidase and antioxidative enzyme activities

1. Changes in MAO and antioxidative enzymes copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD) and catalase (CAT) activities were examined in the hypothalamus and the hippocampus of Wistar rats exposed to cold stress (6 °C) for 180 min and heat stress (38 °C) for 60 min.

2. Extreme environmental temperatures caused stressor-specific changes in the hypothalamic and hippocampal MAO and antioxidative enzyme activities, being dependent on the stressor applied (cold or heat) but not on the brain region studied (the hypothalamus or hippocampus).

Effect of prolonged cold exposure on monoamine oxidase activity and kinetics and on serotonin metabolism in the rat brain

Effects of long-term cold exposure on the content of serotonin and its metabolite 5-hydroxyindolacetic acid (5-HIAA) and monoamine oxidase (MAO) activity and kinetic parameters (Km and Vmax) of oxidative deamination of serotonin in rat brain stem. The increase of 5-HIAA level in the initial period of chronic cold exposure was determined by the blockade of active metabolite transport from the brain. The level of serotonin and the rate of its catalytic deamination by MAO were found to be decreased in cold-adapted rats. The magnitude of the Km of serotonin deamination was unchanged.     

Twenty-four hour rhythms in serum and brain indoleamine concentrations: tryptophan-5-hydroxylase and monoamine oxidase activity in the rat.

The relationship between the 24 h rhythm in 5-hydroxy-tryptamine (5HT) levels in rat brain, the availability of precursors of 5HT and the concentration of its major metabolite, 5-hydroxyindole acetic acid (5HIAA) has been investigated. Serum total and "free" tryptophan (TRY) levels and brain TRY levels all show a 24 h rhythm with highest concentrations in the middle of the dark phase i.e. 12 h displaced from that of the 5HT rhythm. No 24 h variation in either tryptophan-5-hydroxylase or monoamine oxidase activity was detected, nor did brain 5-hydroxytryptophan (5HTP) levels vary with clock hour. Changes in 5HIAA concentration paralleled those of 5Ht. The uptake of 14C-5HTP, 14C-TRY and 14C-5HT into homogenates of the septal region of rat brain did not display a circadian rhythm, although there was evidence that uptake of 14C-TRY in an isolated synaptosomal preparation from the same region was greater during the light phase, indicating the possibility that uptake of the precursor into the nerve ending may be, in part, responsible for the 24 h rhythm in brain 5HT. It is concluded that brain 5HT levels are independent of the serum or brain TRY concentrations measured. Since changes in 5HT with clock hour are paralleled by changes in 5HIAA, it also seems unlikely that the increase in brain 5HT during the light phase is caused by a decreased release of 5HT from nerve endings.