Phosphorylation and the reduced nicotinamide adenine dinucleotide oxidase reaction in Streptococcus agalactiae.
Cell-free extracts from aerobically grown Streptococcus agalactiae cells possess a reduced nicotinamide adenine dinucleotide (NADH) oxidase which is linked to oxygen. It is inhibited by cyanide, although cytochromes evidently are not involved. Adenosine triphosphate (ATP) formation occurs during the reaction, but 66 to 75% of the total ATP is formed nonoxidatively. The remaining 25 to 35% of the ATP formation is related to the oxidation of NADH. The formation of ATP in the oxidative reaction can be prevented by excluding oxygen or adding cyanide to prevent NADH oxidation. It can also be prevented by adding methylene blue or pyruvate, which bypasses electron transport to oxygen, but does not interfere with NADH oxidation. Potential sources of ATP, such as glycolysis, the pyruvate oxidase reaction, or the oxidative pentose cycle, are not present, and the high nonoxidative ATP formation is ascribed to the adenylate kinase reaction. The reaction requires adenosine diphosphate (ADP) as a phosphate acceptor. NADH oxidation is independent of ADP. Antimycin A, amytal, and 2,4-dinitrophenol decreased, but did not prevent, oxidative formation of ATP. P:O ratios ranged from 0.15 to 0.25. All of the oxidative activity was in the soluble portion of the cell-free extracts.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000096:biosynthesis* / Q000378:metabolism; D000654:Amobarbital / Q000494:pharmacology; D000968:Antimycin A / Q000494:pharmacology; D002474:Cell-Free System; D003486:Cyanides / Q000494:pharmacology; D004140:Dinitrophenols / Q000494:pharmacology; D008365:Manometry; D008751:Methylene Blue / Q000494:pharmacology; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation; D010088:Oxidoreductases / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology; D010759:Phosphorus Isotopes; D010770:Phosphotransferases / Q000378:metabolism; D011773:Pyruvates / Q000494:pharmacology; D013291:Streptococcus / Q000201:enzymology / Q000378:metabolism*; D014461:Ultracentrifugation
D003486:Cyanides; D004140:Dinitrophenols; D010759:Phosphorus Isotopes; D011773:Pyruvates; D009243:NAD; D000968:Antimycin A; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D010770:Phosphotransferases; D000251:Adenosine Triphosphatases; D000654:Amobarbital; D010100:Oxygen; D008751:Methylene Blue