A possible physiological role of taurine in the adult female rat liver.

In agreement with previous results (Awapara, 1956), we noted that the taurine level in the liver of the adult female rat is higher than in the adult male rat: 9.44 and 2.08, respectively, expressed as a concentration (mumoles/g of liver). Furthermore, we observed a significant decrease of the taurine level (expressed similarly) in the liver of the lactating rat: 1.84 twenty-one days after the birth of pups. These observations suggest a physiological role for the higher concentration of taurine in the liver of the adult female rat: in other words, a reserve of taurine, whatever its origin in the adult female rat, may be needed for pup development.

Synthesis of taurine in rat liver and brain in vivo.

The in vivo formation of taurine and the analysis of labeled taurine precursors was examined in rat brain and liver at different times after an intracisternal injection of [35S]cysteine and an intraperitoneal injection of [3H]cysteine, simultaneously administered. The distribution pattern of radioactivity was similar in liver and brain. Most of the labeling in both organs (85% in brain and 80% in liver) was recovered in glutathione (oxidized and reduced), cysteic acid, cysteine sulfinic acid, hypotaurine, cystathionine, and a mixed disulfide of cysteine and glutathione. The relative rates of labeling of cysteine sulfinic acid and taurine in liver and brain suggest than in vivo, liver possesses a higher capacity for taurine synthesis than brain. A small amount of [3H]taurine was detected in brain after intra peritoneal injection of [3H]cysteine. The time of appearance of this [3H]taurine as well as the fact that it occurs when [3H]cysteine is not detectable in brain or plasma suggests that it was probably not synthesized in brain from labeled precursors but formed elsewhere and transported into the brain through an exchange process.

Effect of dietary cholic acid and cholesterol on liver and kidney cystathionase and cysteine sulfinate decarboxylase activities and taurine concentrations in the rat.

Hepatic cystathionase and cysteine sulfinate decarboxylase activities are drastically affected by cholic acid added to the diet without cholesterol. When cholic acid and cholesterol are given together, only cysteine sulfinate decarboxylase activity is changed. Neither kidney enzyme activity nor taurine concentrations in the liver and kidney are noticeably modified, whatever the diet.

Dietary casein levels and taurine supplementation. Effects on cysteine dioxygenase and cysteine sulfinate decarboxylase activities and tuarine concentration in brain, liver and kidney of the rat.

Activities of cysteine dioxygenase (CO) and cysteine sulfinate decarboxylase (CSD) and the concentrations of taurine (T) in brain, liver and kidney of rats fed on diets containing 18% casein (A), 60% casein (B) and 17% casein supplemented with 1% of taurine (+T), were measured. Regardless of the diet, the three measurements were the same in the brains of the animals in the three groups. In the liver and the kidney, CO activity was also the same in all three diets, but a decrease of CSD activity associated to an increase of T was observed in rats fed on diet B. The taurine-supplemented diet led to an increase in T concentration.

Cysteine oxidase and cysteine sulfinic acid decarboxylase in developing rat liver.

The patterns of development of cysteine oxidase (CO) and cysteine sulfinic acid decarboxylase (CSD) in rat liver are not similar. It was observed that CO is not under sex control as CSD is. The results obtained agree with the idea that, in liver, as well as in brain, CSD is the limiting factor for the regulation of taurine biosynthesis.

Cystathionine in rat brain: catabolism in vivo.

The content of cystathionine was measured in 35 rat brains; the range was 10-120 nmol/g wet weight and thus the variability of cystathionine content in rat brain was emphasized. The regional distribution of cystathionine was also determined: the highest level was found in cerebellum; the lowest level was observed in the white and gray matter of the hemispheres. These results are different from those obtained in other species. The radioactive metabolites formed from L-(35S)cystathionine injected intracisternally were measured in brains of rats killed at the following times after injection: 0.25, 1, 2, 4,6, 9, 16, and 27 hr. The radioactivity was found both in the proteins and in the acid-soluble fraction. In the acid-soluble fraction the radioactivity was found in various ninhydrin-reacting compounds: [cysteic cysteine sulfinic] acid, taurine, reduced and oxidized glutathione, cystine, cystathionine, and a compound tentatively identified as the mixed disulfide of cysteine and glutathione. The radioactivity of cystathionine decreased exponentially between the 1st and the 27th hour after injection and its half-life was estimated to be about 5 hr. The radioactivity in the other ninhydrin-reacting compounds increased until the 9th hour after injection, then decreased. Half of this radioactivity was present in reduced glutathione, the rest being shared equally between: [cysteic cysteine sulfinic] acid, taurine, and the mixed disulfide. It is worthwhile to note that the radioactivity in the cystine fraction was always very low.

Regional and subcellular distribution of taurine-synthesizing enzymes in the rat central nervous system.

The distribution of cysteine oxidase (CO) and cysteine sulfinate decarboxylase (CSD) was examined in 12 regions of the rat central nervous system (CNS). The distribution of CO activity, expressed as µmol of cysteine sulfinate formed per h per g, was the following: hypothalamus, superior and inferior colliculi, 94-99 µmol/h/g; olfactory bulbs, cerebral cortex, striatum, and hippocampus, 44-51 µmol/h/g; cerebellum, 71 µmol/h/g; pons-medula and spinal cord, 94 and 60 µmol/h/g, respectively. The distribution of CSD activity expressed as µmol of cysteine sulfinate decarboxylated per h per g was the following: hypothalamus and colliculi, 14-21 µmol/h/g; olfactory bulbs, cerebral cortex, striatum, hippocampus, and cerebellum, 8-13 µmol/h/g; pons-medulla, 7.3; and spinal cord, 3.6 µmol/h/g. No CSD activity was detected in sciatic nerve. The subcellular distribution of CO and CSD activities was studied in hypothalamus, colliculi, and cerebral cortex. CO activity was localized in synaptosomes, mitochondria, and microsomes. CSD was primarily confined to the crude mitochondrial fraction and after subfraction, recovered mainly in the synaptosomal fraction.

Free amino acids and related substances in human glial tumours and in fetal brain: comparison with normal adult brain.

An ion exchange automatic chromatographic analysis of the free amino acid concentrations of 18 human glial tumours and of 4 human fetal brains was carried out and the concentrations were compared to those of 13 biopsy specimens of normal adult brain. In addition, the concentrations of the amino acids of the glial tumours were compared to those of 7 intracerebral metastases of various origin. The chromatograms of several tumour specimens showed an unidentified peak overlapping proline. As far as the amino acid concentrations are concerned they varied depending upon the origin of the sample. The concentrations of most amino acids were higher in fetal brain than in adult brain with the exception of aspartic acid, glutamic acid, glutamine, cystathionine and GABA. Two peptides: glutathione and homocarnosine were absent in fetal brain and were present in adult brain. In glial tumours, homocarnosine and some amino acids, namely aspartic acid, glutamic acid and GABA, showed lower concentrations than in normal brain. Some amino acids were in the same concentration as in normal brain: taurine, phosphoethanolamine, glutamine and cystathionine. Most of the others were in higher concentrations than in normal brain, mainly proline. The results suggest that the concentrations of 5 compounds: taurine, proline, cystathionine, GABA and homocarnosine, taken as a whole, provide information on the origin of the sample.

New insights into the active center of rat liver cystathionase.

Treatment by urea of purified rat liver cystathionase (L-Cystathionine cysteine-lyase (deaminating), EC 4.4.1.1) provoked a similar alteration of two activities of the enzyme, namely cysteine desulfhydration and homoserine deamination. Since the decreases of the two activities were also comparable as a result of chymotrypsin digestion of the enzyme, these observations suggest that the two sites responsible for the one and the other activites are in close proximity. Studies of the effect of derivatives of substrates (S-carboxymethylcyste-ine, S-carboxyethylcysteine, S-carboxymethylhomocysteine and S-carboxyethylhomocysteine) on both activities were performed. All of them inhibited cysteine desulfhydration and homoserine deamination; in several cases, the type of inhibition was also determined. The results are in agreement with the hypothesis that each of the two sites of the active center has, at least, three binding points which "recognise" groupings of substrates or of inhibitors, and this led us to propose a model for the active center. Each site has an -NH-2 binding point, hence the active center has two -NH-2 binding points; therefore, as cystathionase consists of four subunits and contains four molecules of pyriodoxal phosphate, it might be of interest to determine whether the smallest active molecule is the dimer.

Rat liver cysteine sulfinate decarboxylase: purification, new appraisal of the molecular weight and determination of catalytic properties.

Rat liver cystein sulfinate decarboxylase (L-cystein sulfinate carboxylase) was purified approximately 500-fold. By cellulose acetate and polyacrylamide gel electrophoresis or by analytical ultracentrifugation, the purified enzyme appears to be nearly homogeneous. The Stokes radius (3.4 nm) and sedimentation coefficient (6.5 S) were determined. The molecular weight, calculated and experimentally estimated is around 100 000 and the enzyme is constituted of two identical subunits whose molecular weights are 55 000. The role of pyridoxal phosphate as coenzyme was demonstrated and the requirement for free sulhydryl groups for activity was studied. The ability of native pure cysteine sulfinate decarboxylase to also decarboxylate cysteate was stressed: therefore, we concluded that in rat liver a single protein catalyzed both reactions, although only the decarboxylation of cysteine sulfinate is of physiological interest.