title: Purification of cytochrome c oxidase by lysine-affinity chromatography.
abstract: A method for the purification of cytochrome c oxidase that is based on the affinity of this enzyme for polycations such as poly-L-lysine is described. When detergent extracts of bovine cardiac mitochondria were applied to either a poly-L-lysine-agarose or a lysine-Sepharose column at low ionic strength, cytochrome c oxidase was found to adhere tightly, whereas the bulk of the proteins were eluted by washing with the same buffer. The cytochrome c oxidase was eluted by application of a linear potassium chloride gradient to the columns. The resulting enzyme was identical to that obtained by more traditional purification methods in terms of its subunit composition, optical and resonance Raman spectra, and cytochrome c oxidizing activity. When detergent extracts of spheroplasts from Paracoccus denitrificans were applied to these columns, the cytochrome c oxidase from this organism was also found to adhere tightly. Thus this purification method appears applicable to both prokaryotic and eukaryotic forms of the enzyme. The advantages of this new purification method are that it is less labor intensive than the traditional procedure and less expensive than methods based on cytochrome c-affinity chromatography.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.
authors.forename: J
authors.initials: J
authors.lastname: Felsch
authors.forename: S
authors.initials: S
authors.lastname: Kotake
authors.forename: R A
authors.initials: RA
authors.lastname: Copeland
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification; D002417:Cattle; D002846:Chromatography, Affinity; D003576:Electron Transport Complex IV / Q000302:isolation & purification*; D008929:Mitochondria, Heart / Q000201:enzymology; D010231:Paracoccus denitrificans / Q000201:enzymology; D011107:Polylysine; D013104:Spheroplasts / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D011107:Polylysine; D003576:Electron Transport Complex IV