title: Spectroscopic determination of cytochrome c oxidase content in tissues containing myoglobin or hemoglobin.
abstract: A simple spectroscopic method for determining the cytochrome c oxidase, cytochrome a, a3, content in tissue and mitochondria samples independent of myoglobin or blood contamination is described. Using tissue homogenates solubilized in Triton X-100, this assay relies on the selective reduction of mitochondrial cytochromes by the action of potassium cyanide. Monitoring the optical absorbance of these samples at 605 nm provided a quantitative determination of cytochrome c oxidase content in the presence of myoglobin or blood. The cytochrome c oxidase content of porcine heart mitochondria and whole tissue was determined to be 0.85 nmol/mg protein and 30.5 nmol/g wet wt, respectively.
authors.affiliation: National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: R S
authors.initials: RS
authors.lastname: Balaban
authors.forename: V K
authors.initials: VK
authors.lastname: Mootha
authors.forename: A
authors.initials: A
authors.lastname: Arai
mesh_terms: D000818:Animals; D003902:Detergents; D003576:Electron Transport Complex IV / Q000032:analysis*; D005069:Evaluation Studies as Topic; D006454:Hemoglobins / Q000032:analysis*; D008929:Mitochondria, Heart / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D009211:Myoglobin / Q000032:analysis*; D017830:Octoxynol; D011190:Potassium Cyanide; D013053:Spectrophotometry / Q000379:methods*; D013552:Swine
chemical_list: D003902:Detergents; D006454:Hemoglobins; D009211:Myoglobin; D017830:Octoxynol; D003576:Electron Transport Complex IV; D011190:Potassium Cyanide