Deficiencies in Complex I subunits of the respiratory chain in Parkinson's disease

Purification and amino acid sequencing of NAF, a novel neutrophil-activating factor produced by monocytes☆

Separation of mammalian cytochrome c oxidase into 13 polypeptides by a sodium dodecyl sulfate-gel electrophoretic procedure.

The mitochondrial cytopathies are a heterogeneous group of diseases associated with defects in mitochondrial ATP production that affect the brain, skeletal muscle, heart, kidney, and liver. Mitochondria produce ATP by oxidative phosphorylation (OXPHOS). OXPHOS involves approximately 100 polypeptides, most of which are encoded in the Mendelian-inherited nuclear genes, but 13 of which are encoded in the maternally-inherited mitochondrial DNA (mtDNA).

Oxidative phosphorylation diseases. Disorders of two genomes.

Cytochrome c oxidase deficient fibres in the limb muscle and diaphragm of man without muscular disease: An age-related alteration

Publisher Summary This chapter discusses the procedure for the isolation of cytochrome- c oxidase from mitochondria of different vertebrate tissues, involving extraction of matrix proteins and most other membrane proteins with nonionic detergents, chromatography on DEAE-cellulose in the presence of Triton X-100, and ammonium sulfate fractionation in the presence of sodium cholate. Cytochrome- c oxidase, the terminal enzyme of the respiratory chain of most aerobic organisms, is a membrane-bound protein complex composed of a variable number of subunits, depending on the evolutionary stage of the organism. Tissue-specific isozymes of mammalian cytochrome- c oxidase are detected, based on differences in (1) apparent molecular weight after SDS-gel electrophoresis, (2) N-terminal amino acid sequence, (3) immunological reactivity of nuclear encoded subunits, (4) kinetic properties, and (5) reactivity of carboxylic groups in the presence and absence of cytochrome- c . The mitochondrially encoded subunits of cytochrome- c oxidase from different tissues of the same organism are suggested to be identical. The methods for the identification of cytochrome-c oxidase isozymes always include separation of the 13 subunits by SDS-gel electrophoresis, representing the most difficult step, as no other method is available for the complete separation of all subunits in one step.

Isozymes of cytochrome-c oxidase: characterization and isolation from different tissues.

Immunological and chemical characterization of rat liver cytochrome c oxidase

With a high-resolving dodecyl sulfate electrophoretic system rat liver cytochrome c oxidase was separated into 13 different polypeptides. An antiserum against rat liver holocytochrome c oxidase immunoreacted with all 13 polypeptides, as demonstrated by immunofluorescence after transfer of the separated Coomassie blue-stained bands on nitrocellulose and coupling with FITC-protein A ("western blot"). Polypeptide-specific antisera reacted only with their corresponding polypeptides indicating that the various protein bands are represented by individual polypeptides. From total proteins of rat liver, kidney, heart, spleen and skeletal muscle mitochondria, only the cytochrome c oxidase polypeptides showed immunofluorescence with an antiserum against the rat liver holoenzyme. In contrast to the polypeptide from liver, polypeptide VIa from heart and skeletal muscle showed little or no reactivity, indicating a tissue-specificity of this polypeptide. Mitochondrial proteins from pig, bovine and blackbird heart were incubated with an antiserum against the rat liver holoenzyme. Immunoreaction was found with most cytochrome c oxidase polypeptides but not with polypeptide VIa. This result demonstrates less immunological relationship between tissue-specific polypeptides (VIa, VIIa and VIII) of the same species than between tissue-unspecific polypeptides of different species.

Tissue-specificity overrides species-specificity in cytoplasmic cytochrome c oxidase polypeptides.

Isolated rat liver cytochrome c oxidase was cross-linked with the cleavable reagents dimethyl-3,3′-dithiobispropionimidate (DTBP), 3,3′-dithiobis(succinimidyl)propionate (DSP) and cupric di(1,10-phenanthroline) (CuP). The cross-linked products were separated by high-resolving two-dimensional dodecyl sulfate gel electrophoresis, which separates all thirteen polypeptides of the mammalian enzyme. With cupric di(1,10-phenanthroline) seven polypeptides (I–III, Va, Vb, VIIb and VIII) were cross-linked with each other and with itself, indicating the occurrence of free –SH groups in these polypeptides and a rearrangement of the native structure of the complex by cupric di(1,10-phenanthroline). With dimethyl-3,3′-dithiobispropionimidate or 3,3′-dithiobis(succinimidyl)propionate all nuclear-coded polypeptides, with the exception of polypeptide VIIa, formed cross-linked products with the three ‘catalytic’ polypeptides I–III, which are coded on mitochondrial DNA. Five additional cross-linked pairs were found between nuclear coded polypeptides. The close arrangement of nuclear coded polypeptides with the catalytic polypeptides suggests a regulatory function of these polypeptides.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1985.tb08641.x

Jochen Jarausch

Papers

Separation of mammalian cytochrome c oxidase into 13 polypeptides by a sodium dodecyl sulfate-gel electrophoretic procedure.

Isozymes of cytochrome-c oxidase: characterization and isolation from different tissues.

Immunological and chemical characterization of rat liver cytochrome c oxidase

Tissue-specificity overrides species-specificity in cytoplasmic cytochrome c oxidase polypeptides.

Structure of the cytochrome c oxidase complex of rat liver