Quantitative determination of cytochromes in the aerobic respiratory chain of Escherichia coli by high-performance liquid chromatography and its application to analysis of mitochondrial cytochromes.

Abstract

A method was established for quantitative determination of the composition and the amounts of cytochromes in the aerobic respiratory chain of Escherichia coli. Cytochromes were solubilized with 3%(W/V) Sarkosyl and the extract was analyzed by high-performance liquid chromatography on a gel permeation column of TSK gel-G3000SW. This analytical system required only about 0.5 mg of membrane protein and 20 min per assay. The effects of the gene dosage of F-prime factors and plasmids on cytochromes were analyzed with this system, and the system was applied to the analysis of mitochondrial cytochromes.