Quinolinate synthetase: the oxygen-sensitive site of de novo NAD(P)+ biosynthesis.
The ability of niacin to relieve the growth-inhibiting effect of hyperoxia on Escherichia coli can be attributed to the dioxygen sensitivity of quinolinate synthetase. The activity of this enzyme within E. coli was diminished by exposure of the cells to 4.2 atm O2, while the activity in extracts was rapidly decreased by 0.2 atm O2. Neither catalase nor superoxide dismutase afforded detectable protection against the inactivating effect of O2, indicating that H2O2 and O2- were not significant intermediates in this process. Nevertheless, H2O2 at 1.0 mM did inactivate quinolinate synthetase, even under anaerobic conditions and in the absence of catalatic activity which might have generated O2. Addition of paraquat to aerobic cultures of E. coli caused an inactivation of quinolinate synthetase, which may be explained in terms of an increase in the production of H2O2. The O2-dependent inactivation of quinolinate synthetase in extracts was gradually reversed during anaerobic incubation and this reactivation was blocked by alpha, alpha'-dipyridyl or by 1,10-phenanthroline. The sequence of the quinolinate synthetase "A" protein contains a--cys-w-x-cys-y-z-cys--sequence, which is characteristic of (Fe-S)4-containing proteins. This sequence, together with the effect of the Fe(II)-chelating agents, suggests that the O2-sensitive site of quinolinate synthetase is an iron-sulfur cluster which is essential for the dehydration reaction catalyzed by the A protein.
Gardner
P R
PR
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Fridovich
I
I
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D000693:Anaerobiosis; D004926:Escherichia coli / Q000201:enzymology; D005296:Ferrous Compounds / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007506:Iron-Sulfur Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology*; D010269:Paraquat / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D013481:Superoxides / Q000494:pharmacology
D005296:Ferrous Compounds; D007506:Iron-Sulfur Proteins; D009097:Multienzyme Complexes; D010618:Phenanthrolines; D013481:Superoxides; C022512:quinolinic acid synthetase; D015082:2,2'-Dipyridyl; D006861:Hydrogen Peroxide; D010269:Paraquat; D010100:Oxygen; C025205:1,10-phenanthroline