Tryptophan Pyrrolase Activity Research Articles

The Relationship Between Tryptophan Pyrrolase Activity and mRNA Expression in Rats Following Chronic Ethanol Administration and Withdrawal

Ethanol exerts a variety of effects on tryptophan and 5-hydroxytryptamine (5-HT) metabolism in man and in experimental animals. In rat models. whose tryptophan metabolism is nearest to that of man, chronic ethanol administration enhances brain 5-HT synthesis whereas inhibition of synthesis occurs during the subsequent withdrawal period. Both effects are caused by corresponding changes in tryptophan availability to the brain and are thought to be due to inhibition and enhancement of liver tryptophan pyrrolase activity respectively. It has been suggested that ethanol acts chronically by inhibiting apo-tryptophan pyrrolase synthesis. We have studied the relationship between tryptophan pyrrolase activity and the expression of hepatic tryptophan pyrrolase mRNA in rats chronically administered ethanol by means of an ethanol-containing liquid diet. Wistar rats were fed ethanol diet at concentrations of 5% and 8% (v/v) and for duration's of two and four weeks. Rats were withdrawn in a starved state for seven hours by which time there were demonstrable signs of withdrawal. Matched controls were fed equal amounts of isocaloric ethanol-free liquid diet. The livers were removed from each of the animals and both tryptophan pyrrolase activity and synthesis were measured. Tryptophan pyrrolase activity was decreased before withdrawal and greatly increased over baseline following withdrawal.

Requirement for tryptophan by milkfish (Chanos chanos Forsskal) juveniles

Groups of milkfish juveniles (mean initial weight 7.7 g) were fed semipurified diets containing 0.9, 1.4, 2.1, 3.1, 4.1 and 6.1 g tryptophan/kg dry diet for 12 weeks. The mean crude protein content of the diets (containing white fishmeal, gelatin and free amino acid mixture to simulate the pattern of hydrolysed milkfish protein) was 49%. On the basis of the growth response, the tryptophan requirement of milkfish juveniles was estimated to be 3.1 g/kg diet. Fish fed low levels of tryptophan exhibited low weight gains and poor feed conversion ratios. Survival (92-100%) was consistently high in all treatments. Fish fed diets containing tryptophan levels greater than 3.1 g/kg had slightly lower survival rates. The activity of hepatic tryptophan pyrrolase showed no significant differences with increasing dietary tryptophan levels. No nutritional deficiency signs were observed other than the depression in growth rates in fish given the tryptophan deficient diets.

Activity and Partial Purification of Liver Tryptophan Pyrrolase in Rainbow Trout.

Compounds that activate tryptophan pyrrolase in other animals like methylene blue, hematin, EDTA, and calciumion did not affect fish enzyme activity. Neutralized ascorbic acid showed an inhibitory effect. Time-activity and enzyme concentration-activity curves were determined in three methods of assay. Km was estimated via the Lineweaver-Burk plot to be 80mM. The enzyme was partially purified by a four-step process: solubilization, DEAE Sephadex A-50 chromatography (batch method), and by two consecutive DEAE Sepharose CL-6B chromatographs. The partially purified enzyme was highly unstable so that no characterization was at all possible.

Control of cytochromes P450 expression in Gunn rat liver: Implication of the intracellular heme pool

The absence of changes in the overall hepatic cytochrome P450 content after administration of 3-methylcholanthrene (MC) to congenitally jaundiced Gunn rats is believed to be related to a limited heme availability in this strain of rat. The amount of available heme, estimated by tryptophan pyrrolase activity, shows a substantial decrease in control Gunn versus control Wistar rats. This reduction is moderately enhanced by MC treatment in Gunn rats but is abolished after phenobarbital administration. Heme oxygenase activity is diminished in Gunn rats and consequently is not responsible for the decrease in the hepatic heme availability. These data point out that the depletion of the intracellular heme can lead to a limitation in the synthesis of cytochrome P450 isoenzymes in the MC-induced Gunn rat.

The effects of lofepramine and desmethylimipramine on tryptophan metabolism and disposition in the rat

Acute and chronic administration of lofepramine and its major metabolite desmethylimipramine (DMI) to rats elevates brain tryptophan concentration, thereby enhancing cerebral 5-hydroxytryptamine (5-HT) synthesis, by increasing the availability of circulating tryptophan to the brain, secondarily to inhibition of liver tryptophan pyrrolase (tryptophan 2,3-dioxygenase, l-tryptophan:O 2 oxidoreductase, decyclizing; EC 1.13.11.11) activity. The pyrrolase inhibition by lofepramine occurs independently of metabolism to DMI, because it can be demonstrated directly in vitro. Lofepramine also differs from DMI in its action profile on the above and related aspects of tryptophan metabolism and disposition. These results demonstrate that lofepramine influences tryptophan and 5-HT metabolism and disposition independently of its major metabolite DMI, and are discussed briefly in relation to the mechanism of action of antidepressants.

In vitro effect of dimethoate on the activity of tryptophan pyrrolase in rat liver

Total and holo-enzyme activities of tryptophan 2,3-dioxygenase were measured in vitro in the presence and absence of the organophosphorous insecticide, dimethoate. Addition of dimethoate to the reaction mixture decreased the activities of both total and holo-forms. Total and holo-enzyme activities were decreased by 34% and 26%, respectively, by 1 mM dimethoate. On the other hand, 5 mM dimethoate resulted in 56% and 34% inhibition to total and holo-enzyme activities, respectively. Lineweaver-Burk plot of the total-enzyme activity at different tryptophan concentration in the presence of 2 mM dimethoate gave uncompetitive type of inhibition.

Effects of acute paroxetine administration on tryptophan metabolism and disposition in the rat

1 The effects of acute oral administration of paroxetine on tryptophan metabolism and disposition were examined in the rat. 2 Basal liver tryptophan pyrrolase activity was inhibited by paroxetine in vitro and after oral administration. Maximum inhibition was caused by a 1 mg kg-1 dose. 3 Paroxetine administration also inhibited pyrrolase activity that had previously been enhanced by hormonal induction by cortisol or cofactor activation by haematin. The cortisol induction of the enzyme was, however, not inhibited by pretreatment of rats with paroxetine. 4 Paroxetine increased tryptophan availability to the brain, because of the above pyrrolase-inhibitory mechanism. Cerebral 5-hydroxytryptamine (5-HT) synthesis was accordingly enhanced, though this was apparent only with doses of the drug of up to 1 mg kg-1. With larger doses, decreased 5-HT turnover, probably as a result of 5-HT uptake inhibition, was the more dominant feature. 5 Paroxetine lowered circulating corticosterone concentration, but did not influence those of albumin, non-esterified fatty acids or glucose. 6 It is concluded that, in addition to inhibiting brain 5-HT turnover, paroxetine also, in common with 20 other antidepressants, enhances 5-HT synthesis by increasing brain tryptophan concentration secondarily to inhibition of liver tryptophan pyrrolase activity.

The effect of space flight on the board of the satellite cosmos 2044 on plasma hormone levels and liver enzyme activities of rats

The aim of present experiment was to study the changes of corticosterone, insulin and glucose levels in plasma, of the activity of enzymes involved in aminoacid metabolism in liver and the binding of insulin to specific receptors of cell membrane from liver and also of adipose tissue of rats exposed to space flight for 14 days on biosatellite Cosmos 2044. Adult male Wistar rats (body mass 300–370 g) were divided into five groups: intact control rats (AC), rats exposed to space flight (F), animals in synchronous model experiment (S), rats in antiorthostatic hypokinesia (A) and so called operated control group (C). Half of all groups (5 animals) except the intact control were operated 3 days before the experiment (fibulas on both hind legs were broken). The flight animals were sacrificed 5–6 hours after landing. It was observed that plasma insulin levels are increased in rat exposed to 14-day space flight and in synchron experiments. A significant increase of plasma glucose levels was found in flight rats in spite of high insulin concentrations suggesting that in rats exposed to 14-day space a deterioration of tissue sensitivity to insulin could by present. No significant differences of specific insulin binding to liver plasma membrane fraction in flight and intact control animals were observed. A decrease of insulin binding capacity in liver was found in rats in antiorthostatic hypokinesia (A). However in the membrane of adipocytes an important increase of insulin receptors was noted in rats subjected to space flight. These results suggest, that the liver and adipocyte insulin receptors of flight rats did not respond to the increased plasma insulin levels by “down regulation”. The determination of plasma corticosterone levels showed that in flight rats and in animals exposed to antiorthostatic hypokinesia the plasma hormone levels are significantly elevated. A significant increase of tyrosine aminotransferase and tryptophan pyrrolase activities in liver of flight rats and those exposed to hypokinesia was observed. Also the elevation of alanine aminotransferase in liver was observed in flight rats, while, the activity of aspartate aminotransferase in liver was similar in control and flight animals. These results showed that the changes in liver enzyme activities in rats after 14-day space flight are in agreement with the results observed in previous experiments after a shorter space flight (7 days).

Metabolism of lysine and tryptophan in growing rats at various dietary protein levels.

The metabolic fates of the carbon skeletons of L-[U-14C]lysine and L-[methylene-14C]tryptophan were investigated in growing rats fed with diets containing different percentages of protein calories (0, 5, 10, 15, and 30PC%) at 4100 kcal of metabolizable energy per kg of diet. The incorporation of 14C into the body protein 12 hr after the injection of 14C-labeled lysine or tryptophan was extremely high in the 0 to 10PC% groups (about 80% of the injected dose being recovered), and the value decreased in the higher PC% groups. The expired 14CO2 production from both labeled amino acids was depressed in the lower PC% groups, and then increased with increasing levels of dietary protein, showing an inverse pattern to that of the 14C incorporation into the body protein. The activities of hepatic lysine-ketoglutarate reductase and tryptophan pyrrolase were increased with increasing dietary protein levels, and the response pattern of these enzyme activities resembled that of the expired 14CO2 production from these 14C-amino acids. These results indicate that the metabolic responses of lysine and tryptophan to dietary protein levels change at around 10PC%, where the growth rate reached its approximate maximum.

Kinetic properties of human liver tryptophan pyrrolase.

Human liver tryptophan pyrrolase (TPO) activity exhibited substrate level regulation. TPO showed biphasic activity to tryptophan when low ascorbate was used as an activator. The high affinity form (Km for tryptophan: 0.05 mM) was promoted by low ascorbate and low tryptophan. The low affinity form (Km for tryptophan: 0.4 mM) was induced by high concentrations of tryptophan or ascorbate. Both high and low affinity forms showed the same affinity to oxygen. The high affinity form was also induced by pyrroloquinoline quinone, but this effect was decreased by catalase, suggesting the participation of H2O2.

In vivo evidence that insulin does not inhibit hepatic tryptophan pyrrolase activity in rats

Previous reports have indicated that insulin administration triggers an early increase in plasma tryptophan (TRP) levels in fasted rats. Then, the present study was undertaken to investigate the putative role of liver tryptophan pyrrolase (TPO) in this short-term effect of insulin. In 24 hr fasted rats, doses of insulin that triggered an increase in plasma TRP levels (i.e., 2–3 I.U./kg, 1 hr) did not alter either holoenzyme or total enzyme activity. In another series of experiments, the administration of insulin (2 I.U./kg) to 24 hr fasted rats promoted biphasic time effects on plasma TRP levels and liver TPO activity. Thus, insulin initially triggered a rise in plasma TRP (without any change in liver TPO activity) and then increased liver TPO activity whilst plasma TRP returned toward control levels. In addition, hypercorticosteronemia was evidenced throughout the first phasis. Lastly, the influence of insulin administration (2 I.U./kg) on fasting-induced TPO induction was analysed. Whereas fasting increased liver TPO activity in a time-dependent manner, insulin administration (2 I.U./kg, 30 min) did not modify either plasma TRP or liver TPO activity. The data reported herein bring evidence that the effect of insulin administration on circulating TRP is not mediated by an inhibition of hepatic TPO.

Injected tryptophan increases brain but not plasma tryptophan levels more in ethanol treated rats

In previous studies, long term treatment with ethanol has been shown to enhance brain 5-hydroxytryptamine 5-(HT) metabolism by increasing the activity of the regulatory enzyme tryptophan hydroxylase and or availability of circulating tryptophan secondarily to an inhibition of hepatic tryptophan pyrrolase. In the present study ethanol treatment given for two weeks decreased hepatic apo-tryptophan pyrrolase but not total tryptophan pyrrolase activity in rats. Tryptophan levels in plasma and brain did not increase significantly. But there was a marked increase of 5-HT but not 5-hydroxyindoleacetic acid (5-HIAA) concentration in brain, suggesting a possible increase in the activity of tryptophan hydroxylase. The effect of a tryptophan load on brain 5-HT metabolism was therefore compared in controls and ethanol treated rats. One hour after tryptophan injection (50mg/kg i.p.) plasma concentrations of total and free tryptophan were identical in controls and ethanol treated rats, but the increases of brain tryptophan 5-HT and 5-HIAA were considerably greater in the latter group. The results are consistent with long term ethanol treatment enhancing brain serotonin metabolism and show that brain uptake/utilization of exogenous tryptophan is increased in ethanol treated rats and may be useful to understand the role and possible mechanism of tryptophan/serotonin involvement in mood regulation.

Liver tryptophan pyrrolase. A major determinant of the lower brain 5-hydroxytryptamine concentration in alcohol-preferring C57BL mice

The lower brain 5-hydroxytryptamine concentration in alcohol-preferring C57BL, compared with -non-preferring CBA, mice is caused by a decrease in circulating tryptophan availability to the brain secondarily to a higher liver tryptophan pyrrolase activity associated with a higher circulating corticosterone concentration. Activity or expression of liver tryptophan pyrrolase and/or their induction by glucocorticoids may be important biological determinants of predisposition to alcohol consumption.

Heme precursor 5-aminolevulinic acid alters brain tryptophan and serotonin levels without changing pineal serotonin and melatonin concentrations.

The daytime administration of the heme precursor 5-aminolevulinate (5-ALA) has been shown to reduce brain tryptophan and serotonin levels owing to saturation of liver tryptophan pyrrolase. Saturation of this enzyme with heme results in enhanced activity, leading to increased catabolism of tryptophan and thus making less tryptophan available to the brain. Tryptophan is also the precursor of melatonin, a primary secretory product of the pineal gland. Reduced melatonin levels, which could be a consequence of altered tryptophan metabolism, have been associated with depressive disorders in humans. In view of this, in the present study we examined the effects of 5-ALA administration on forebrain tryptophan and serotonin levels as well as pineal serotonin, melatonin, and the pineal melatonin-forming enzymes N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT). 5-ALA induced a reduction in forebrain tryptophan and serotonin levels during the light phase and caused the opposite effect in the dark phase. Allopurinol, an inhibitor of hepatic tryptophan pyrrolase activity, prevented the reduction in the indole levels induced by 5-ALA. Although 5-ALA induced these changes in the forebrain, it did not alter either daytime or nighttime pineal NAT, HIMOT, serotonin, or melatonin levels. It appears that high serotonin levels and melatonin production in the pineal are conserved even in the face of low circulating tryptophan levels, changes which alter brain tryptophan and serotonin concentrations.

Contribution of methylsulfonyl metabolites of m-dichlorobenzene to the heme metabolic enzyme induction by the parent compound in rat liver

In the present study, we investigated the contribution of methylsulfonyl metabolites derived from m-dichlorobenzene ( m-DCB) on the heme metabolic enzyme induction by the parent compound in rats. The time courses of the effects of a single ip administration of m-DCB (200 mg/kg, 1.36 mmol/kg) and 2,4- and 3,5-dichlorophenyl methyl sulfones (2,4- and 3,5-DCPSO 2Mes) (each 50 μmol/kg) on hepatic microsomal cytochrome P450 content were almost in parallel with those on the total heme content in liver microsomes. m-DCB significantly increased the heme oxygenase activity, but 2,4- and 3,5-DCPSO 2Mes did not. On the other hand, m-DCB and both methyl sulfones markedly enhanced the δ-aminolevulinic acid (ALA) synthetase activity. No change was observed in percentage saturation of the tryptophan pyrrolase activity after administration of m-DCB, whereas this ratio at 6 hr after injection of 3,5-DCPSO 2Me was increased. In the liver of the dl-buthionine-( S,R)-sulfoximine (BSO)-treated rats dosed with m-DCB, both of 2,4- and 3,5-DCPSO 2Mes were present at significantly lower concentrations than in non-BSO-treated rats. Additionally, the m-DCB did not elevate the ALA synthetase activity in the BSO-treated rat. On the other hand, the administration of either 2,4- or 3,5-DCPSO 2Mes to BSO-treated rats resulted in induction of ALA synthetase. m-DCB and 2,4- and 3,5-DCPSO 2Mes produced a dose-related increase in liver levels of methyl sulfones. The changes in the ALA synthetase activity after the administration of varying doses of m-DCB were similar to those after the administration of 2,4- or 3,5-DCPSO 2Mes, whereas the sum of the concentration of two methyl sulfones in the liver of rats dosed with m-DCB was almost the same as the concentration of methyl sulfone after the administration of either 2,4- or 3,5-DCPSO 2Mes. The results strongly suggest that the methyl sulfones derived from m-DCB, i.e., 2,4- and 3,5-DCPSO 2Mes, contribute highly to the induction of the ALA synthetase activity by the parent compound.

Depletion of liver regulatory heme in benzene exposed rats

The effect of a single dose of benzene (0.5 ml/kg body wt i.p.) on the heme saturation of tryptophan pyrrolase activity in liver was examined. There was a significant decrease in the heme saturation of hepatic tryptophan pyrrolase, suggesting depletion of “regulatory heme”. After benzene administration there was significant increase in σ-aminolevulinate (ALA) synthetase activity (approx. 2-fold) while σ-aminolevulinate dehydratase activity was significantly decreased, however, ferrochelatase and heme oxygenase activities were unaltered. Administration of tryptophan to benzene pretreated rats showed a reversal of benzene effects on heme synthesizing enzymes: there is an increase in the heme saturation of tryptophan pyrrolase and decrease in σ-aminolevulinate synthetase. However, there was no significant alteration in the activity of σ-aminolevulinate dehydratase.

Effects of the haem precursor 5-aminolaevulinate on tryptophan metabolism and disposition in the rat.

5-Aminolaevulinate administration to rats inhibits cerebral 5-hydroxytryptamine synthesis by decreasing tryptophan availability to the brain secondarily to activation of hepatic tryptophan pyrrolase. The results show that tryptophan metabolism and disposition can be influenced by changes in liver haem concentration, and are discussed briefly in relation to mood disorders in the hepatic porphyrias.

Inhibition of rat Liver Tryptophan Pyrrolase Activity and Elevation of Brain Tryptophan Concentration by Administration of DL-α-Amino-β-Pyridinepropanoic Acid (Pyridylalanine) Analogs

Single doses of DL-alpha-amino-beta-(2-pyridine)propanoic acid (2-PA, 100 mg/kg) significantly decreased the holoenzyme and apoenzyme activities of rat liver tryptophan pyrrolase (TP) and increased brain tryptophan, serotonin (5-HT) and 5-hydroxyindole-3-ylacetic acid concentrations. 2-PA had no inhibitory effect on either of the enzyme activities in vitro, but its expected metabolites were effective. Single doses of DL-alpha-amino-beta-(3-pyridine)propanoic acid (3-PA, 100 mg/kg) decreased only the holoenzyme activity and elevated brain tryptophan and its metabolites levels in rats. 3-PA and its metabolite, 3-pyridylpyruvate, inhibited only the holoenzyme activity in vitro. DL-alpha-Amino-beta-(4-pyridine)propanoic acid (4-PA) caused significant changes in liver TP (holo- and apoenzyme forms) activity and brain tryptophan concentration only after repeated administration (100 mg/kg/day). 4-PA was a weak inhibitor of the holoenzyme, but its metabolites apparently inhibited the holo- and apoenzyme activities in vitro. These findings suggest that PA analogs (and/or their metabolites) increased brain tryptophan (and hence 5-HT synthesis) by directly inhibiting liver TP activity.

Effects of primaquine on hepatic microsomal haemoproteins and drug oxidation

Administration of the antimalarial agent primaquine to male rats (50 mg/kg i.p. daily for 4 days) resulted in a 30% decrease in hepatic microsomal cytochrome P-450 ( P-450) content; levels of other microsomal haemoproteins were unaltered. Kinetic analysis of 2 mixed-function oxidase activities (aminopyrine N-demethylase and aniline p-hydroxylase) in primaquine-pretreated rat liver microsomes revealed a significant and similar decrease in the maximal reaction velocities of these enzymes ( V max), but the apparent Michaelis constants ( K m) were not changed. The activity of mitochondrial δ-amonolaevulinic acid synthetase (the rate-liniting step in haem biosynthesis) was normal in primaquine-pretreated rat liver but haem oxygenase activity (the rate-limiting step in haem degradation) was elevated approximately 2-fold. Haem availability for haemoprotein assembly (determined as the haem-saturaiton ratio of the cytosolic haemoprotein tryptophan pyroolase) was also normal although the absolute activity of tryptophan pyrrolase was decreased after primaquine pretreatment. order to cacilitate an analysis of the P-450 isozyme profile in control and primaquine-treated rat live,, total microsomal P-450 was isolated by hydrophobic affinity chromatography on n-octylamino-Sepharose 4B of stained polyacrylamide gels following electrophoresis of these partially-purified P-450 fractions indicated that primaquine exposure did not selectively decrease any of the 3 protein bands in the P-450 molecular weight region (48–56 kD). These observations, when considered together, suggest that primaquine may affect P-450 and mixed-function oxidase activity by inhibition of protein synthesis. The characteristic rapid turnover rates of P-450 isozymes may predispose these haemoproteins to the toxic effects of primaquine whereas those haemoproteins that turn over less rapidly, such as cytochrome b 5, appear to be less susceptible. Microsomal haem oxygenase activity may be elevated after primaquine administration since lowered haemoprotein requirements for haem could result in excess haem levels within the hepatocyte.

Effect of sodium copper chlorophyllin on lipid peroxidation. VIII. Its effect on carbon tetrachloride-induced liver injury in rats.

The present study was carried out to investigate the effectiveness of sodium copper chlorophyllin (Cu-Chl-Na) as an in vivo lipid antioxidant by examining its effect on CCl4-induced hepatic injury. Rats received two i.p. injections of Cu-Chl-Na (50 or 100mg/kg) in and 2h prior to CCl4 administration (1 ml/kg, p.o.). It was found that in the pretreated rats, there was effective depression of both the CCl4-induced lipid peroxidation in hepatic microsomes and the CCl4-induced increase of lipid peroxides in whole liver at 0.5 and 24h after the dosing of CCl4, respectively. Cu-Chl-Na pretreatment also clearly inhibited the elevation of serum levels of glutamic oxaloacetic and glutamic pyruvic transaminases and isocitric dehydrogenase, and the increase in liver triglyceride level caused by CCl4. In addition, the hepatotoxin-induced decrease of hepatic tryptophan pyrrolase activity was substantially prevented in the rats pretreated with Cu-Chl-Na. Since the CCl4-induced peroxidation of liver lipids is generally accepted to be involved in the pathogenesis of its hepatotoxicity, the above in vivo antioxidative effect of Cu-Chl-Na is considered to play an important role in its protection of rats against the liver damage produced by CCl4.