Apoprotein formation and heme reconstitution of cytochrome P-450cam.
Apoprotein suitable for heme reconstitution has been prepared by an acid/butanone extraction of cytochrome P-450cam at pH 2.5. Absorption spectra of apo-P-450cam indicate less than 2% residual holoenzyme. Four tryptophan residues per molecule were estimated from the aromatic absorbance region of denatured apoprotein. Heme-reconstituted holoprotein was purified in 30% yield to a specific activity equivalent to the native enzyme. Absorption and EPR spectra of 57Fe- and 54Fe-heme-enriched P-450cam reveal complete restoration of the native active site.
D001059:Apoproteins; D055598:Chemical Phenomena; D002621:Chemistry; D003577:Cytochrome P-450 Enzyme System; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme; D011549:Pseudomonas / Q000201:enzymology; D013057:Spectrum Analysis
D001059:Apoproteins; D006418:Heme; D003577:Cytochrome P-450 Enzyme System