Further observations on the Mg-protoporphyrin IX monomethyl ester (oxidative) cyclase system.
The Mg-protoporphyrin IX monomethyl ester (oxidative) cyclase was strongly inhibited by CN- and N3- in a reconstituted system, but was inhibited slightly or not at all by the same reagents in intact developing chloroplasts. Known inhibitors of cytochrome P-450 processes showed no consistent effect. Benzoquinone and quinol, which can give rise to the same semiquinone by one-electron redox events, were strong inhibitors of the cyclase. It was previously shown that O2 and a source of electrons are required in the cyclization process. The substrates for the dehydrogenases of the pentose phosphate pathway (glucose 6-phosphate and 6-phosphogluconate) were effective reductants in the reconstituted system with supernatant that had been dialysed or passed through Sephadex G-50, in the absence of added NADP+. However, inhibitor studies suggested that the electrons from these sugar phosphates reached the cyclase system via NADPH. Therefore we infer the presence of protein-bound NADP+ that can be reduced by glucose 6-phosphate and 6-phosphogluconate and donate reducing equivalents to the cyclase system. This bound NADPH pool may be particularly effective in the cyclization process, owing to channeling. These findings are discussed in relation to the results of a companion paper [Whyte and Castelfranco (1993) Biochem. J. 290, 361-367] on the breakdown of chloroplast pigments in the same reconstituted system.
Whyte
B J
BJ
Department of Botany, University of California, Davis 95616.
Castelfranco
P A
PA
D000641:Ammonia / Q000494:pharmacology; D003486:Cyanides / Q000494:pharmacology; D065607:Cytochrome P-450 Enzyme Inhibitors; D004583:Electrons; D005958:Glucosephosphates / Q000494:pharmacology; D009243:NAD / Q000737:chemistry; D009249:NADP / Q000737:chemistry; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D011524:Protoporphyrins / Q000737:chemistry*; D014675:Vegetables
D003486:Cyanides; D065607:Cytochrome P-450 Enzyme Inhibitors; D005958:Glucosephosphates; D011524:Protoporphyrins; D009243:NAD; C033434:protoporphyrin IX monomethyl ester; D009249:NADP; D000641:Ammonia; D010105:Oxygenases; C053161:magnesium protoporphyrin monomethyl ester oxidative cyclase