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B Kadenbach

Publications and source records attributed to B Kadenbach.

At least 109 records · Page 6Linked to original sources
PubMed1988

Immunocytochemical studies of cytochrome oxidase subunits in skeletal muscle of patients with partial cytochrome oxidase deficiencies.

Muscle biopsies from 17 patients with partial cytochrome oxidase deficiencies were investigated using immunocytochemical techniques for the localisation of cytochrome oxidase subunits. Antisera to subunits II/III (mitochondrially coded) and subunits IV, Vab, VIbc, VIIa, VIIbc and VIII (nuclear coded) showed clear particulate immunoreactivity in the muscle fibres of normal control biopsies. In the patients studied, muscle fibres with absent or decreased cytochrome oxidase activity also showed decreased immunoreactivity affecting all enzyme subunits. Particularly close correlation was seen between percentages of fibres showing absent enzyme activity and those showing decreased immunoreactivity for subunits II/III which are catalytic in function. The regulatory subunits IV-VIII were affected to varying degrees with different patterns of subunit loss occurring in individual muscle fibres.

Adolescent↗
PubMed1988

Cytochrome oxidase deficiency: immunological studies of skeletal muscle mitochondrial fractions.

We report a 2-year-old girl who presented with delayed development, weakness and persistent vomiting. She had a demyelinating peripheral neuropathy. The activity of cytochrome oxidase in skeletal muscle from the patient was 10% of controls. Immunochemical studies using antibodies to holo-cytochrome oxidase and the individual subunits showed a low concentration of all detectable subunits.

Child, Preschool↗
PubMed1988

Organization and nucleotide sequence of two chromosomal genes for rat cytochrome c oxidase subunit VIc: a structural and a processed gene.

A rat genomic library was screened with a cDNA probe coding for rat liver cytochrome c oxidase subunit VIc (COX-VIc). Out of 16 clones mapped, four inserts were different and did not overlap. Two inserts that gave strong hybridization signals were characterized by sequence analysis. These data show that we have cloned two different genes for rat COX-VIc. The rat COX-VIc-2 gene contains four exons and three introns. The 5' boundary of the first exon was mapped by primer extension experiments. The nucleotide sequence of the first three exons is identical to the sequence of the rat liver cDNA probe; thus, we can conclude that this gene is expressed in rat liver. The second gene (COX-VIc-1) is 88% homologous to the rat liver cDNA and contains an open reading frame of 228 nucleotides capable of coding for a full-length COX-VIc polypeptide of 76 amino acids. This predicted translation product is 79% homologous to the rat liver peptide. The absence of introns and other factors, such as the presence of a 13-bp direct repeat and a poly(A) addition signal, indicate that this locus is a processed gene which may have evolved from spliced mRNA intermediates.

Amino Acid Sequence↗
PubMed1988

Evolution of cytochrome c oxidase.

The current view on the regulatory function of nuclear-encoded subunits of cytochrome c oxidase from higher evolved organisms is presented. The activity of monomeric laurylmaltoside-dissolved, but not of reconstituted cytochrome c oxidase, is strongly affected by anions accompanied by a conformational change of the enzyme, as shown by changed visible spectra. Addition of uncoupler to proteoliposomes induces the same anion sensitivity as obtained with the soluble enzyme, suggesting dissociation of the dimeric membrane-bound enzyme by uncoupler. Nucleotides are suggested to regulate cytochrome c oxidase activity at 3 different sites: 1) Interaction of ATP with a cytosolic site (outside) increases the Km for cytochrome c in the enzyme from bovine heart and Paracoccus denitrificans; 2) binding of ADP at a matrix site decreases, and 3) binding of ATP at another matrix site increases the Km for cytochrome c of the mammalian enzyme.

Amino Acid Sequence↗
PubMed1987

Molecular cloning and further characterization of cDNAs for rat nuclear-encoded cytochrome c oxidase subunits VIc and VIII.

A cDNA library was constructed from poly(A)-rich RNA of H35 rat hepatoma cells by insertion into lambda gt11. Screening with an antiserum to rat liver holocytochrome-c oxidase yielded fifteen different recombinant clones. Eight clones were identified using monospecific antisera to individual subunits of the rat liver enzyme. The cDNA clones coding for subunits VIc and VIII were further characterized by DNA sequence analysis after subcloning in pUC8 and M13 mp9. The deduced amino acid sequences show 80% and 60% homology to the corresponding bovine heart subunits, respectively. From the nucleotide sequences we can conclude that subunit VIc is not synthesized as a larger precursor molecule like most other mitochondrial proteins. The size of the mRNAs coding for subunits VIc and VIII is about 450 nucleotides, as revealed by Northern blot analysis with RNAs from different tissues. The clones were further used as probes for Southern blotting. Restricted high-molecular-mass DNA showed a complex pattern of bands indicating multigene families for both subunits in the rat genome.

Amino Acid Sequence↗
PubMed1987

ADP increases the affinity for cytochrome c by interaction with the matrix side of bovine heart cytochrome c oxidase.

The effect of intraliposomal ADP and ATP on the kinetics of cytochrome c oxidation in reconstituted bovine heart cytochrome c oxidase was measured by the photometric and polarographic method: 1. Intraliposomal ADP decreases and intraliposomal ATP increases the Km for cytochrome c when measured by the photometric assay under uncoupled conditions. 2. The above described effects are not obtained when the kinetics are measured with the polarographic assay. 3. Extraliposomal ATP increases the Km for cytochrome c similar to intraliposomal ATP, but this effect is measured with both methods of assay. 4. Under coupled conditions only a small decrease of the Km for cytochrome c by intraliposomal ADP is found.

Adenosine Diphosphate↗
PubMed1987

The cytochrome c oxidase from the yeast Candida parapsilosis.

In the yeast Candida parapsilosis, the proteins encoded by mitochondrial DNA are different in number and size from those of Saccharomyces cerevisiae. Nevertheless, the purified cytochrome c oxidase from Candida parapsilosis shows kinetic properties similar to those of Saccharomyces cerevisiae.

Candida↗
PubMed1986

Influence of buffer composition, membrane lipids and proteases on the kinetics of reconstituted cytochrome-c oxidase from bovine liver and heart.

Isolated cytochrome-c oxidases from bovine heart and liver were reconstituted in liposomes with asolectin and the kinetics of cytochrome c oxidation were measured under various uncoupled conditions. With 40 mM KCl, 10 mM Hepes, pH 7.4, the liver enzyme showed a higher Vmax in the polarographic but a lower Vmax in the photometric assay. With 125 mM phosphate buffer at pH 6.0 both enzymes revealed identical kinetics. Reconstitution with pure phosphatidylcholine leads to a low activity, which is specifically stimulated for the heart enzyme by inclusion of 10% cardiolipin. Proteoliposomes of both enzymes prepared with asolectin have a high activity, which is unaffected by cardiolipin. Exchanging the intraliposomal buffer, Hepes, for phosphate causes an opposite change of the Vmax and a similar change of the Km for both enzymes suggesting a conformational change of the extraliposomal binding domain for cytochrome c through the membrane. Proteases change the kinetics of both enzymes, but to a different degree. The data indicate a complex and tissue-specific influence of nucleus-coded subunits on the catalytic activity of cytochrome-c-oxidase.

Animals↗
PubMed1986

Analysis of the Cu, Fe, and Zn contents in cytochrome C oxidases from different species and tissues by proton-induced X-ray emission (PIXE).

The Cu, Fe and Zn contents in isolated cytochrome c oxidase preparations from heart, liver, diaphragm or kidney of bovine, pig and rat was measured by proton-induced X-ray emission (PIXE). The average Cu/2Fe ratio was 2.73 and Zn/2Fe ratio 0.98. Correspondingly a Cu/Zn ratio of 2.76 was found. Dialysis of the bovine heart enzyme against increasing EDTA concentrations up to 30 mM did not change this result. It is concluded that all isozymes of mammalian cytochrome c oxidase contain 3 Cu, 2 Fe and 1 Zn per monomeric catalytic unit.

Animals↗
PubMed1986

Specific effects of ATP on the kinetics of reconstituted bovine heart cytochrome-c oxidase.

Bovine heart cytochrome-c oxidase was reconstituted in liposomes and the kinetics of cytochrome c oxidation were measured by the polarographic and photometric method under uncoupled conditions in the presence of various polyvalent anions. In order to distinguish between specific and unspecific ionic effects of ATP, the photolabelling reagent 8-azido-ATP was applied. Covalently bound ATP at the enzyme complex caused the same increase of Km for cytochrome c as free ATP, if measured by the photometric assay. The increase of Km by photolabelling with 8-azido-ATP was completely prevented by ATP, but not by ADP. The data indicate the occurrence of a specific binding site for ATP at the cytosolic side of cytochrome-c oxidase, which, after binding of ATP, changes the kinetics of cytochrome c oxidation.

Adenosine Diphosphate↗
PubMed1986

Tissue-specific and species-specific distribution of -SH groups in cytochrome c oxidase subunits.

Cytochrome c oxidase was isolated from pig, bovine, rat and human tissues including liver, heart, diaphragm and kidney. The native and the sodium-dodecyl-sulfate (SDS)-dissociated enzymes were labelled under optimal conditions with N-ethyl-[2,3-14C]maleimide before and after reduction with dithiothreitol, separated into 13 subunits by SDS gel electrophoresis and the radioactive bands were visualized by fluorography. In some cases the radioactive bands were cut out and counted. All isozymes were labelled in subunits I, III, Va and VIIb, and in subunit II after reduction. Labelling of subunit Vb was equivocal, and in no case were subunits IV and VIc labelled. All other subunits were labelled tissue-specifically and/or species-specifically. No differences were found between labelling of the native and SDS-dissociated enzyme. By relating the molar amount of bound N-ethylmaleimide to the known amount of cysteines in subunits of bovine heart cytochrome c oxidase, the percentage of -SH group reactivity was calculated. Only the cysteine of subunit Va was found to be 100% reactive. The distinct and different reactivity of subunit VIIb as compared to subunits VIIa and VIIc clearly establishes this polypeptide as an independent subunit of mammalian cytochrome c oxidase.

Animals↗
PubMed1986

Isolation and characterization of human heart cytochrome c oxidase.

Cytochrome c oxidase was isolated from human hearts and separated by SDS gel electrophoresis. The identity of polypeptide bands with known subunits was demonstrated by immunoblotting with monospecific antisera to rat liver cytochrome c oxidase subunits. The polarographically determined kinetics of cytochrome c oxidation were similar to those reported for the bovine heart enzyme.

Animals↗
PubMed1986

Regulation of respiration and ATP synthesis in higher organisms: hypothesis.

The present view on the regulation of respiration and ATP synthesis in higher organisms implies only Michaelis-Menten type kinetics and respiratory control as regulatory principles. Recent experimental observations, suggesting further regulatory mechanisms at respiratory chain complexes, are reviewed. A new hypothesis is presented implying regulation of respiration and ATP synthesis in higher organisms mainly via allosteric modification of respiratory chain complexes, in particular of cytochrome c oxidase. The allosteric effectors, e.g., metabolites, cofactors, ions, hormones, and the membrane potential are suggested to change the activity and the coupling degree of cytochrome c oxidase by binding to specific sites at nuclear coded subunits. Recent results on the structure and activity of cytochrome c oxidase, supporting the hypothesis, are reviewed.

Adenosine Triphosphate↗
PubMed1986

Fatal mitochondrial cardiomyopathy in Kearns-Sayre syndrome with deficiency of cytochrome-c-oxidase in cardiac and skeletal muscle. An enzymehistochemical--ultra-immunocytochemical--fine structural study in longterm frozen autopsy tissue.

Morphological studies in a 26-year-old man with long-standing Kearns-Sayre syndrome, with cardiac arrhythmias and a fatal congestive cardiomyopathy, revealed a mitochondrial myopathy of both skeletal and myocardial muscle (Hübner et al. 1986). Histochemical investigation of cytochrome-c-oxidase showed multiple enzyme defects of both cardiac and skeletal muscle present in myocytes with normal and abnormal numbers of mitochondria demonstrated by ultracytochemistry. Immunohistochemical studies with antibodies against the holoenzyme and various subunits revealed that in the heart the enzyme defect affected both contractile and conductive fibres and was characterized by a severe reduction but not a complete loss of nuclear and mitochondrially coded immunoreactive enzyme protein. In skeletal muscle, however, where up to 30% of the fibres lacked enzyme activity, immunoreactivity was reduced only very occasionally. These results are most consistent with a defective enzyme assembly in the inner mitochondrial membrane and probably indicate heterogeneity of mitochondria, i.e. organ-specific pathological reaction patterns.

Adult↗
PubMed1985

Orientation of rat liver cytochrome c oxidase subunits investigated with subunit-specific antisera.

The orientation of rat liver cytochrome c oxidase subunits in the inner mitochondrial membrane was investigated with monospecific antisera against subunit II and nine nuclear-coded subunits. Mitoplasts were incubated with the antisera and the amount of bound antibodies was determined either directly with fluorescein-conjugated protein A or indirectly by back-titration of unbound antibodies with a nitrocellulose immunoassay. All subunits were found oriented to the cytosolic side, except subunits VIb and VIIc which did not react with their corresponding antisera. Antisera against subunits I, III and Vb were not available.

Animals↗