Preparation and characterization of S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]glutathione and its derivatives as proposed precursors of S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]cysteine, a compound found in human urine.

Formation of 3-[(carboxymethyl)thio]-3-(1H-imidazol-4-yl)propanoic acid (I) and S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]cysteine (II), compounds found in human urine, has been demonstrated by enzymatic degradation of S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]glutathione (III). Compound (III) was chemically synthesized in 72% yield by incubating the reaction mixture of trans-urocanic acid and 3-fold excess GSH at 65 degrees C for 1 wk, which was accompanied by formation of N-(S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]cysteinyl)glycine (IV) in 15% yield. S-[2-Carboxy-1-(1H-imidazol-4-yl)ethyl]-N-gamma-glutamylcysteine (V) was produced by partial hydrolysis of compound (III) in HCl. The synthesized compounds were characterized mainly by fast-atom bombardment mass spectrometry and high-voltage paper electrophoresis as well as chemical degradation. Incubation of compound (III) with rat kidney homogenate in a Tris buffer (pH 8), formed compound (II) in 80% yield possibly via compound (IV). Yield of compound (II) was increased by adding glycylglycine to the reaction mixture. However, little degradation of compound (III) occurred in the use of rat liver, brain, heart or spleen homogenate as the enzyme source. Compound (II) was further metabolized to compound (I) by incubation with rat kidney homogenate in a phosphate buffer of pH 7.4. From these results, we suggest that the urinary compounds are products of enzymatic degradation of compound (III) and that GSH may participate in the metabolism of urocanic acid, the first catabolite of L-histidine.

Excretion of sulfate and taurine in rats fed with a high protein diet.

Sulfate and taurine are the main metabolites of L-cysteine in mammals and are excreted in the urine. The effect of a high protein diet on the ratio of sulfate to taurine excretion was studied in rats using synthetic 25% (standard protein diet group, group A) and 40% (high protein diet group, group B) casein diets. Average taurine and sulfate excretions (mumol/kg of body weight per day) were 280.4 +/- 93.8 and 943.2 +/- 144.8 in group A and 553.4 +/- 124.5 and 2675.0 +/- 390.9 in group B, respectively. Thus, the average taurine/sulfate ratio in group A was 0.30 +/- 0.08. By a single administration of 5 mmol of L-cysteine/kg of body weight in group A, the average taurine and sulfate excretions increased to 1127.5 +/- 120.2 and 4043.0 +/- 305.6, respectively, but the taurine/sulfate ratio changed only slightly (0.28). The average taurine/sulfate ratio in group B was 0.22 +/- 0.07, a significantly lower ratio than that in group A, which means that daily intake of a high protein diet resulted in more sulfate excretion. The taurine/sulfate ratio in group B was affected only slightly (0.19) by the cysteine administration as well. These results suggest that the ratio of taurine and sulfate production was determined by dietary protein content and that the increase in sulfate production is larger than that of taurine production when the intake of dietary protein is increased.

Excretion of taurine and sulfate in rats fed with a low protein diet.

The effects of a low protein diet on the excretion of sulfate and taurine, major metabolites of L-cysteine in mammals, were studied in rats fed with synthetic 10% (group A) and 25% (group B) casein diets. The average excretions of total taurine (taurine plus hypotaurine) and total sulfate (free plus ester sulfate) (mumol/kg of body weight per day after the adaptation to the synthetic diet) in group A were 14.2 +/- 13.4 and 122.3 +/- 39.6, respectively, which were very low compared with 280.4 +/- 93.8 and 943.2 +/- 144.8, respectively, in group B. The taurine/sulfate ratio in group A was 0.12 +/- 0.11, which was significantly lower than that (0.30 +/- 0.08) in group B. A single intraperitoneal injection of 5 mmol of L-cysteine per kg of body weight in group A resulted in an increase in average taurine and sulfate excretion to 693.4 +/- 195.6 and 2440.6 +/- 270.0, respectively, and thus the average taurine/sulfate ratio increased to 0.29. These increases were transient and low taurine excretion resumed again 24 h after the L-cysteine administration. L-Cysteine injection in group B resulted in a similar increase in taurine and sulfate excretion, but the ratio changed only slightly (0.28). The present results suggest that in vivo production of taurine is reduced preferentially over sulfate production when sulfur amino acid supply is limited.

Increased excretion of taurine, hypotaurine and sulfate after hypotaurine loading and capacity of hypotaurine metabolism in rats.

Hypotaurine was intraperitoneally injected into rats and urinary taurine, hypotaurine and sulfate were determined. Taurine excretion increased dose-dependently when 1 to 7 mmol of hypotaurine per kg of body weight was administered. The total excretion and the increased excretion (difference of those before and after the loading) were 2328 +/- 219 and 1948 +/- 153 mumol per kg of body weight per day, respectively, at 7 mmol of hypotaurine loading. Hypotaurine excretion was negligible in the normal rat urine. However, it increased when hypotaurine was loaded. Hypotaurine excretion at 7 mmol of hypotaurine loading was 2282 +/- 258 mumol per kg per day. These results indicate that the capacity of hypotaurine oxidation to taurine in rats is more than 2 mmol per kg per day under the present experimental conditions. Sulfate excretion increased significantly when more than 3 mmol of hypotaurine per kg of body weight was injected. When 5 and 7 mmol of hypotaurine was loaded, the increased excretion of sulfate was 619 +/- 205 and 632 +/- 118 mumol per kg per day, respectively. It was confirmed that in vitro incubation of hypotaurine and 2-oxoglutarate with rat liver homogenate results in the formation of L-glutamate and sulfate. Present findings indicate that hypotaurine in vivo was mainly oxidized to taurine and that it was partly metabolized to sulfate via transamination reaction.