dihydroneopterin-triphosphate and sepiapterin

The structure of dyspropterin, a new name given to an intermediate which is formed from dihydroneopterin triphosphate in the biosynthetic pathway of tetrahydrobiopterin, has been studied. Sepiapterin reductase (EC 1.1.1.153) was found to reduce dyspropterin to tetrahydrobiopterin in the presence of NADPH. Several lines of evidence showing the formation of tetrahydrobiopterin have been presented. Stoichiometric analysis revealed that there is a 1:2 relationship between the production of biopterin and the oxidation of NADPH during the reductase-catalyzed reduction of dyspropterin. The tetrahydrobiopterin production from dyspropterin was enhanced by dihydropteridine reductase (EC 1.6.99.7). Dyspropterin could also serve as a cofactor in phenylalanine hydroxylase (EC 1.14.16.1) system. These results are consistent with the view that dyspropterin is 6-(1,2-dioxopropyl)-5,6,7,8-tetrahydropterin. Based on our findings, the biosynthetic pathway of tetrahydrobiopterin from dihydroneopterin triphosphate has been discussed.

Topics: Alcohol Oxidoreductases; Biopterins; Catalysis; Chemical Phenomena; Chemistry; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Dihydropteridine Reductase; Hydroxylation; Neopterin; Oxidation-Reduction; Pteridines; Pterins; Spectrophotometry, Ultraviolet

9 partially purified enzyme (Enzyme A) from Drosophila melanogaster Aatalyzes the conversion of 7,8- dihydroneopterin triphosphate to a compound that, from its ultraviolet absorption spectrum and other characteristics, appears to be 6- pyruvoyl -tetrahydropterin. This product can be converted to 6-lactoyl-tetrahydropterin in the presence of another partially purified enzyme (Enzyme B) and NADPH, and to 5,6,7,8-tetrahydrobiopterin in the presence of a third enzyme preparation (biopterin synthase) and NADPH. The enzymically-produced 6-lactoyl-tetrahydropterin, when exposed to air, is oxidized nonenzymically to sepiapterin (6-lactoyl-7,8- dihydropterin ). The results indicate that although 6-lactoyl-tetrahydropterin can be converted enzymically to tetrahydrobiopterin, neither it nor sepiapterin is an obligate intermediate in the conversion of 7,8- dihydroneopterin triphosphate to tetrahydrobiopterin.

Topics: Animals; Biopterins; Biotransformation; Catalysis; Drosophila melanogaster; Edetic Acid; Magnesium; NADP; Neopterin; Pteridines; Pterins

Using Escherichia coli guanosine triphosphate cyclohydrolase, dihydroneopterin triphosphate was synthesized from guanosine triphosphate and was compared with sepiapterin as a substrate for tetrahydrobiopterin formation in bovine adrenal medulla extracts. The dihydrofolate reductase inhibitor, methotrexate, blocks the formation of tetrahydrobiopterin from sepiapterin but not from dihydroneopterin triphosphate. Reduced nicotinamide adenine dinucleotide phosphate and a divalent metal ion are required in partially purified preparations (gel filtration of 40-60% ammonium sulfate fraction on Ultrogel ACA-34) for the biosynthesis of tetrahydrobiopterin from dihydroneopterin triphosphate. Sepiapterin was converted only to dihydrobiopterin in the same fractions since dihydrofolate reductase was removed. The evidence indicates that both dihydroneopterin triphosphate and sepiapterin are good precursors of tetrahydrobiopterin but they are not on the same pathway, contrary to previous proposals.

Topics: Adrenal Medulla; Animals; Biopterins; Cattle; Chemical Phenomena; Chemistry; Guanosine Triphosphate; Methotrexate; Neopterin; Pteridines; Pterins

The enzymatic conversion of the D-erythro-dihydroneopterin triphosphate [H2-neopterin-(P)3] to sepiapterin occurs via a nonphosphorylated intermediate as shown by others. We have developed a high-performance liquid chromatography assay for this intermediate and have found that the intermediate (X) and two related compounds (X1 and X2) can be formed nonenzymatically under certain conditions from H2-neopterin-(P)3. The reaction is catalyzed by tris(hydroxymethyl)aminomethane, dependent upon H2-neopterin-(P)3 concentration, significant at temperatures greater than 80 degrees C, and maximal between pH 8.5 and 9.5 (as determined at 25 degrees C). All three compounds were purified, and it was found that both X and X1 can serve as substrates for the enzymatic, NADPH-dependent synthesis of sepiapterin. From the kinetics of formation from H2-neopterin-(P)3 and the similarity of the ultraviolet spectra, it is clear that X, X1, and X2 are closely related compounds. None of the three compounds is reduced by NaBH4; only X1 is sensitive to periodate oxidation. All three can be oxidized with iodine to give rise to highly fluorescent compounds that in turn can be reduced by NaBH4 to give rise to the respective parent compounds. These latter observations indicate that X, X1, and X2 are dihydropterins. These results are discussed relative to the proposed structures for enzymatically produced X. The methods described for the nonenzymatic synthesis of X and its purification should allow preparation of large amounts of X for future study.

Topics: Drosophila; Hydrogen-Ion Concentration; Magnesium; NADP; Neopterin; Pteridines; Pterins