Purification of all thirteen polypeptides of bovine heart cytochrome c oxidase from one aliquot of enzyme Characterization of bovine fetal heart cytochrome c oxidase

Abstract

A protocol has been worked out for separating all thirteen different polypeptides in the beef heart cytochrome c oxidase complex from a single aliquot of enzyme. This involves an initial separation of polypeptides by gel filtration on a Biogel P-60 column in SDS, a step which purifies subunits C_IV and C_VIII and gives mixtures of C_V+C_VI, ASA, AED and STA, as well as C_VII, C_IX and IHQ. These mixtures are then resolved by reverse-phase high-performance liquid chromatography. The separation procedures have been applied to fetal heart cytochrome c oxidase of gestation between 100 and 200 days. No differences were found in the N-terminal sequences of any of the cytoplasmically made subunits or in the entire sequence of C_IX between late fetal and adult forms of the enzyme.