Biochemical aspects and functional role of the copper-containing amine oxidases

The copper-containing amine oxidases of the class EC 1.4.3.6 share many biochemical similarities. They contain cupric copper and catalyse the same general reaction. In mammals, diamine oxidase has a role in the metabolism of histamine, some other diamines and spermine oxidase, involved in the metabolism of polyamines. However, the functional role of benzylamine oxidase (BAO) and the tissue semicarbazide-sensitive amine oxidases (SSAO) is still under investigation. Circulating BAO is derived from the tissue SSAO. It has a high affinity for benzylamine and its role in the extra-cellular matrix includes the maturation of pro-elastin and control of the inflammatory process.     

Contribution of different amine oxidases to the metabolism of dopamine in bovine retina.

The contribution of monoamine oxidase (MAO) A, MAO B and semicarbazide-sensitive amine oxidase (SSAO) to the metabolism of dopamine in the bovine retina was studied. These activities were present in the optic nerve, iris, choroid and bovine retina, but they were absent in the lens. SSAO activity towards dopamine was present in the choroid and the retina, but not in the iris or the optic nerve. The corresponding kinetic values for this substrate in the retina and the choroid showed higher affinity for MAO A (Km 271 and 197 microM, respectively) than for MAO B (Km 861 and 404 microM, respectively). This effect was counteracted by the higher Vmax value for MAO B resulting in the Vmax/Km ratio being similar for both cases. The absence of detectable SSAO activity towards dopamine in these last two tissues contrasts with the presence of that enzyme when benzylamine was studied as substrate. These results indicate that two different SSAO activities could be present in the bovine eye.     

Some new functions of amine oxidases

Two contrasting topics are examined in this account: the protective actions of amine oxidases (AOs) resulting from the elimination and/or modulation of the levels of polyamines and some biogenic amines, such as histamine, in anaphylactic shock and the cell damaging effect of AOs catabolic products. Other functions of the plasma copper-containing amine oxidase are considered; namely the modification of some proteins by oxidation of their free amino groups, the auto-regulation of the catalytic activity of AOs, the protective effect against free radicals, and the regulation of K+ - channels.     

Guanabenz as inhibitor of copper-containing amine oxidases

The effects of guanabenz on some copper containing amine oxidases are described. Guanabenz 'in vitro' inhibits pig plasma benzylamine oxidase with a IC50 M 5.1 +/- 0.8 X 10(-6) M. It also inhibits pig kidney diamine oxidase and rat liver mitochondrial monoamine oxidase at higher concentrations. The significance of this property of guanabenz is discussed.     

Inter-ethnic differences in xenobiotic metabolism

The aim of safety assurance procedures is to determine a level of intake (the acceptable daily intake (ADI) or tolerable daily intake (TDI)) that is without adverse health effects in the human population. The majority of studies on inter-ethnic differences in xenobiotic metabolism have concentrated in the incidence of expression of the poor metaboliser phenotype for a number of drug metabolising enzymes. Such ethnic differences can result in different incidences of individuals at higher risk, but this would not affect the safety assurance/risk assessment outcome unless poor metaboliser status was not recognised in the database used for the initial assessment and calculation of ADI or TDI. Of far greater importance are ethnic differences which result in population differences in the mean values, and/or the extent of variability within the population, for key kinetic parameters such as the internal dose or area under the plasma concentration time curve (AUC). There are few studies on inter-ethnic differences in sensitivity and most of these relate to in vivo differences for therapeutic/pharmacological agents, so that the reported data reflect both kinetic and dynamic variability. Inter-ethnic differences are limited in extent and well within the uncertainty factor of 10 used for human variability.     

Role of gut in xenobiotic metabolism

The gastrointestinal tract forms the first line of defense in the body against the main load of xenobiotics. The gastrointestinal mucosa has several mechanisms through which the xenobiotics are modified. The monooxygenase activities in most species are relatively low in the mucosa as compared to the liver, but conjugation, for example, via glucuronide formation proceeds efficiently. UDP-glucuronosyltransferase activities can exceed those in the liver. Glutathione S-transferase activity is also high. The biotransformation activities are readily inducible in the mucosa and this is, at least partly, responsible for the oral-aboral gradient seen in enzyme activities. In rainbow trout glutathione S-transferase is, however, significantly higher at the aboral third than in two oral segments, although in rats the intestinal glutathione S-transferase shows a clear oral-aboral gradient. The gradient is independent of the presence of microflora at least in the case of carboxylesterase and glutathione S-transferase. A similar gradient can also be found from the gut lumen, in both germ-free and specific pathogen-free rats. The cells in the middle of the villi appear to be most responsive under the influence of inducers. The readily occurring induction in the mucosa provides a suitable model for studies on biological effects to defined compounds and mixtures.Dedicated to Professor Dr. med. Herbert Remmer on the occasion of this 65th birthday     

Observations on the substrate specificity of amine oxidases

A number of compounds have been tested as substrates of, first, the amine oxidases of rabbit and guinea-pig liver and of goat, pig, horse, and dog plasma and, second, the “diamine oxidase” of pig kidney. Of the three xylylenediamines [di(aminomethyl)benzenes]tested, m-xylylenediamine was found to be a substrate of the liver oxidase. All the plasma oxidases tested acted on both m- and o-xylylenediamine. Both 3- and 4-picolylamine (3- and 4-aminomethylpyridine) were readily oxidized by the liver enzymes; all the plasma oxidases tested oxidized 2-, 3-, and 4-picolylamine. 4-Picolylamine was more rapidly oxidized by horse and dog plasma than any other substrate hitherto examined.     

The role of microsomal mixed function oxidases in the metabolism and mechanism of action of certain organophosphorus insecticides

The influence of microsomal mixed function oxidases (MFO) on the biotransformation of pesticides was investigated in experiments with perfusion of isolated rat livers and in whole animals. The pesticides examined were 3 organophosphorus insecticides (OP): S-propyl-O-phenyl-O-ethylthiophosphate (heterophos), a mixture of O,O-dimethyl-O-cyclohexylthionphosphate and O,S-dimethyl-O-cyclohexylthiolphosphate (cyclophos), and dithiophosphonate. The indices of NADPH-dependent hydroxylating systems measured in the microsomal fraction of rat liver homogenates were the activity of aniline hydroxylase, aminopyrine demethylase and the amount of cytochrome P-450. It was shown that not only did the specific chemical structure of the OP and induction or inhibition of MFO determine OP activation and detoxification (and thus its biological action), but also the route and frequency of exposure could influence these parameters.     

Impact of environmental exposures on ovarian function and role of xenobiotic metabolism during ovotoxicity

The mammalian ovary is a heterogeneous organ and contains oocyte-containing follicles at varying stages of development. The most immature follicular stage, the primordial follicle, comprises the ovarian reserve and is a finite number, defined at the time of birth. Depletion of all follicles within the ovary leads to reproductive senescence, known as menopause. A number of chemical classes can destroy follicles, thus hastening entry into the menopausal state. The ovarian response to chemical exposure can determine the extent of ovotoxicity that occurs. Enzymes capable of bioactivating as well as detoxifying xenobiotics are expressed in the ovary and their impact on ovotoxicity has been partially characterized for trichloroethylene, 7,12-dimethylbenz[a]anthracene, and 4-vinylcyclohexene. This review will discuss those studies, as well as illustrate where knowledge gaps remain for chemicals that have also been established as ovotoxicants.     

Reduction of fat deposition by combined inhibition of monoamine oxidases and semicarbazide-sensitive amine oxidases in obese Zucker rats

Semicarbazide-sensitive amine oxidase (SSAO) and monoamine oxidases (MAO) are highly expressed in adipocytes and generate hydrogen peroxide when activated. Consequently, high concentrations of MAO- or SSAO-substrates acutely stimulate glucose transport and inhibit lipolysis in isolated adipocytes in a hydrogen peroxide-dependent manner. Chronic treatments with MAO and SSAO substrates also increase in vitro adipogenesis and in vivo glucose utilization and fat deposition in diabetic rodents. To further investigate the interplay between amine oxidases, energy balance and fat deposition, prolonged MAO and/or SSAO blockade was performed in obese rats. Pargyline (P, MAO inhibitor), semicarbazide (S, SSAO inhibitor), alone or in combination (P+S), were daily i.p. administered for 3-5 weeks to obese Zucker rats at doses ranging from 20 to 300 micromol/kg. P+S treatments abolished MAO and SSAO activities in any tested tissue. P and S led to a 12-17% reduction of food intake when given in combination but were inactive when given separately. Despite a similar body weight gain reduction in P+S-treated and pair-fed rats, the mitigation of fat deposition was greater in rats receiving both inhibitors. Adipocytes from P+S-treated rats responded as control to insulin but exhibited impaired responses to tyramine, benzylamine or methylamine plus vanadate when considering glucose transport activation or lipolysis inhibition. Although our results did not directly demonstrate that amines are able to spontaneously produce in vivo the insulin-like effects described in vitro, we propose that P+S-induced reduction of fat deposition results from decreased food intake and from impaired MAO- and SSAO-dependent lipogenic and antilipolytic actions of endogenous or alimentary amines.     

Substances inhibiting amine oxidases. III. Synthesis and amine oxidase inhibition of imidazoquinoline derivatives

The activity of copper and FAD dependent amine oxidases was tested with some derivatives of 3H-imidazo[4,5-h]quinoline and its isomers 3H-imidazo[4,5-f]quinoline, the chemistry of which is described in the literature (1), and Ki calculated. The methyl derivative of 3H-imidazo[4,5-f]quinoline was found to activate the copper bovine serum enzyme, but inhibits the FAD mitochondrial enzyme.