The characterization and magnetic properties of the azide and imidazole derivatives of Pseudomonas nitrite reductase.
Optical absorption, mcd, and epr spectroscopy have been used to characterize the azide and imidazole derivatives of oxidized Pseudomonas nitrite reductase. At pH 7.0 azide binds solely to heme d1 with an affinity constant, Kaff = 360 M-1, whereas imidazole binds to both hemes c and d1 with kaff = 35 and 55 M-1, respectively. Low-temperature mcd and epr spectroscopy indicate that c and d1 are low-spin ferrihemes in both derivatives, although the epr of the heme d1-azide component is very weak and requires explanation. Attempts to obtain a high-spin heme d1 in the intact enzyme using the weak field ligands fluoride and thiocyanate have proved unsuccessful. Electron paramagnetic resonance experiments involving an oxidized enzyme derivatives in which heme d1 is complexed by NO, and hence epr silent, have enabled unambiguous assignment of the epr spectrum of Pseudomonas nitrite reductase.
D001386:Azides; D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme; D007093:Imidazoles; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009572:Nitrite Reductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013053:Spectrophotometry
D001386:Azides; D007093:Imidazoles; D006418:Heme; D009247:NADH, NADPH Oxidoreductases; D009572:Nitrite Reductases