The effect of azide on the spectral and catalytic properties of ascorbate oxidase.
(1) 45% of the total copper of green zucchini ascorbate oxidase is EPR-detectable. At least two species of copper are present, one with a small A parallel (Type 1) and one with a large A parallel (Type 2). Computer simulated spectra indicated 50% contribution by each type of copper. (2) Azide inhibited ascorbate oxidase activity by an uncompetitive mechanism. EPR and optical spectra performed on titration of ascorbate oxidase with azide indicated the formation of a copper-azide complex. The Type 2 copper appears to be the binding site of azide. The involvement of the EPR non-detectable copper as an anion binding site with high affinity toward azide can not be excluded.
D001204:Ascorbate Oxidase / Q000378:metabolism; D001386:Azides / Q000494:pharmacology*; D001665:Binding Sites; D003300:Copper / Q000032:analysis; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D010088:Oxidoreductases; D010944:Plants / Q000201:enzymology; D011485:Protein Binding; D011487:Protein Conformation; D013045:Species Specificity; D013053:Spectrophotometry; D013056:Spectrophotometry, Ultraviolet
D001386:Azides; D003300:Copper; D010088:Oxidoreductases; D001204:Ascorbate Oxidase