Purification and properties of the enzymes from Drosophila melanogaster that catalyze the conversion of dihydroneopterin triphosphate to the pyrimidodiazepine precursor of the drosopterins.
The enzyme system responsible for the conversion of 2-amino-4-oxo-6-(D-erythro-1',2',3'-trihydroxypropyl)-7,8-dihyd roptridine triphosphate (dihydroneopterin triphosphate or H2-NTP) to 2-amino-4-oxo-6-acetyl-7,8-dihydro-3H,9H-pyrimido[4,5-b]-[1,4]diazepine (pyrimidodiazepine or PDA), a precursor to the red eye pigments, he drosopterins, has been purified from the heads of Drosophila melanogaster. The PDA-synthesizing system consists of two components, a heat-stable enzyme and a heat-labile enzyme. The heat-stable enzyme can be replaced by sepiapterin synthase A, a previously purified enzyme required for the Mg2+-dependent conversion of H2-NTP to an unstable compound that appears to be 6-pyruvoyltetrahydropterin (pyruvoyl-H4-pterin). The heat-labile enzyme, purified to near-homogeneity and termed PDA synthase (Mr = 48,000), catalyzes the conversion of pyruvoyl-H4-pterin to PDA in a reaction requiring the presence of reduced glutathione. Because PDA is two electrons more reduced than pyruvoyl-H4-pterin, the reducing power required for this transformation is probably supplied by glutathione. The PDA-synthesizing system requires the presence of another thiol-containing compound such as 2-mercaptoethanol when incubation conditions 2-mercaptoethanol is no longer required. Evidence is presented to indicate that the Drosophila eye color mutant, sepia, is missing PDA synthase.
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification*; D000818:Animals; D001381:Azepines / Q000096:biosynthesis*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D005127:Eye Color; D007658:Ketone Oxidoreductases / Q000302:isolation & purification; D009154:Mutation; D019798:Neopterin / Q000031:analogs & derivatives; D019761:Phosphorus-Oxygen Lyases; D011621:Pteridines / Q000096:biosynthesis* / Q000378:metabolism*; D011622:Pterins; D013053:Spectrophotometry
D001381:Azepines; D011621:Pteridines; D011622:Pterins; C013490:dihydroneopterin triphosphate; C020306:drosopterin; D019798:Neopterin; C032200:2-amino-4-oxo-6-acetyl-7,8-dihydro-3H,9H-pyrimidodiazepine; D000429:Alcohol Oxidoreductases; D007658:Ketone Oxidoreductases; C043498:pyrimidodiazepine synthase; D019761:Phosphorus-Oxygen Lyases; C043497:6-pyruvoyltetrahydropterin synthase