A comparison of the physical and chemical properties of four cytochromes c from Azotobacter vinelandii.
1. A modified method for the separation and purification of four cytochromes c from Azotobacter vinelandii is described. Two new cytochromes c have been purified and are designated cytochromes c(551) and c(555). 2. Additional evidence is presented to establish the dihaem nature of cytochrome c(4). Ultracentrifugation data indicated similar molecular weights for the native and the denatured protein. Cleavage with CNBr yielded seven peptides; the amino acid compositions of the purified peptides were determined. Only one haem peptide was recovered. 3. Cytochromes c(551) and c(555) were characterized as acidic proteins of molecular weights about 12000. The spectral properties, isoelectric points, ;maps' of peptides from CNBr cleavage and amino acid compositions were determined for these two proteins. 4. The spectral properties, isoelectric points, molecular weights, CNBr peptide ;maps', amino acid compositions, relative oxidation-reduction potentials and e.p.r. (electron-paramagnetic-resonance) spectra of the four cytochromes c were compared. Cytochrome c(4) and cytochrome c(551) appear to be distinct proteins. The distinction between cytochromes c(5) and c(555) was not as clear, and our data are inadequate to establish firmly that they are distinct proteins. 5. The dihaem nature of cytochrome c(4) is evident in its e.p.r. spectrum. The e.p.r. spectra are similar to the spectra of mammalian cytochromes c.
D000595:Amino Acid Sequence; D001395:Azotobacter / Q000201:enzymology*; D055598:Chemical Phenomena; D002621:Chemistry; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003488:Cyanogen Bromide; D003574:Cytochrome c Group / Q000032:analysis / Q000302:isolation & purification; D004578:Electron Spin Resonance Spectroscopy; D005292:Ferricyanides; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000032:analysis; D013696:Temperature; D014461:Ultracentrifugation; D014465:Ultrasonics
D003574:Cytochrome c Group; D005292:Ferricyanides; D010446:Peptide Fragments; D003488:Cyanogen Bromide