The heme environment of ovoperoxidase as determined by optical spectroscopy.
Native ovoperoxidase exhibited an optical absorption spectrum with certain similarities to lactoperoxidase, but not horseradish peroxidase, over the pH range 4.5-11.5. Ovoperoxidase had three distinct spectral forms dependent on pH, with transitions at apparent pKa values of 6.6 and 3.0. Complexes of ovoperoxidase with CN-, N3-, F-, or when reduced and ligated to carbon monoxide, CN-, or pyridine, were distinct from other peroxidases. Ovoperoxidase formed two specific and different spectral derivatives at pH 6.0 and 8.0, either in the native state, or when combined with CN-, when reduced, or when reduced and ligated to CN-. The position of the Soret band when mixed with near-stoichiometric amounts of H2O2. This cycling was inhibited by phenylhydrazine, 3-amino-1,2,4-triazole, or low pH (less than or equal to 6). Compound II was formed when ovoperoxidase was mixed with ethyl hydrogen peroxide in a 1:3 ratio, but not with H2O2. With a great excess of H2O2, Compound III was formed at pH 8.0; at pH 6.0 or below, the Soret band shifted slightly with excess of H2O2, but Compound III was never formed. Even when ovoperoxidase was bound to proteoliaisin (Weidman, P. J., and Shapiro, B. M. (1987) J. Cell Biol. 105, 561-567), ovoperoxidase exhibited spectral characteristics of the free enzyme.
Somers
C E
CE
Department of Biochemistry, University of Washington, Seattle 98195.
Shapiro
B M
BM
D000818:Animals; D000838:Anions; D001386:Azides / Q000378:metabolism; D003486:Cyanides / Q000378:metabolism; D005260:Female; D005459:Fluorides / Q000378:metabolism; D037161:Galectins; D006388:Hemagglutinins / Q000235:genetics*; D006418:Heme; D006735:Horseradish Peroxidase; D006863:Hydrogen-Ion Concentration; D007784:Lactoperoxidase; D010063:Ovum / Q000201:enzymology*; D010084:Oxidation-Reduction; D010544:Peroxidases; D012617:Sea Urchins / Q000201:enzymology; D013053:Spectrophotometry
D000838:Anions; D001386:Azides; D003486:Cyanides; D037161:Galectins; D006388:Hemagglutinins; D006418:Heme; D006735:Horseradish Peroxidase; D007784:Lactoperoxidase; D010544:Peroxidases; D005459:Fluorides