Tissue‐specific genes code for polypeptide VIa of bovine liver and heart cytochrome <i>c</i> oxidase

Kadenbach, Bernhard; Hartmann, Renate; Glanville, Robert W.; Buse, Gerhard

doi:10.1016/0014-5793(82)80450-x

Cited by 96 publications

(21 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the concept of isoenzymes for membrane proteins, in particular of mitochondrial membranes, seemed at first bold and premature. it has meanwhile been extended to several other instances such as cytochrome c oxidase [27]. This aspect of organ specificity has also become particularly significant for mitochondrial autoimmune antisera.…”

Section: Discussionmentioning

confidence: 99%

Immunochemical characterization of the adenine nucleotide translocator

Schultheiß

Klingenberg

1984

European Journal of Biochemistry

View full text Add to dashboard Cite

Antibodies have been prepared against purified preparations of the heart and kidney nucleotide translocator in the 'c'-conformation. The results show organ-specific antigenic determinants on the translocator proteins isolated from heart, kidney and liver although a partial cross-reactivity between these three proteins was demonstrable. The organ specificity was observed both with the solubilized and with the membrane-bound translocator protein indicating organ-specific determinants on exposed regions of the carrier. An organ-specific inhibition of the nucleotide transport in heart mitochondria by the heart carboxyatractylate-protein antiserum leads to the conclusion that the organ specificity is at least partially conditioned by the binding site for the substrate and/or the closely linked gate of the carrier protein. Apart from the organ specificity the results also demonstrate a specificity of the antibodies for the translocational conformations of the carrier: the 'c'-conformation stabilized in the carboxyatractylate-protein complex and the 'm'-conformation present in the bongkrekate-protein complex. However, after denaturation of the carboxytraktylate-protein and bongkrekate-protein complexes the binding of the anti-(carboxyatractylate-protein) antiserum to both inhibitor-protein complexes was nearly identical. The conformation specificity was also expressed by the inhibition of the conformation transition from the 'c'-to the .m'-state. This side-specific inhibition of the nucleotide transport and the identical binding activity of the carboxyatractylate-protein antiserum against the denatured carboxyatractylate-protein and bongkrekate-protein complexes suggested that the conformation-specific antigenic determinants are topographic surface regions which are determined by the chain folding.Previous studies indicated that the adenine nucleotide translocator can raise antibodies against antigenic determinants that are strongly conformation dependent. Thus antibodies against the carboxyatractylate-protein complex, representing the 'c'-conformation of the translocator facing the cytosolic side of the inner mitochondria1 membrane, did not react with the bongkrekate-protein complex, representing the 'm'-conformation facing the matrix side of the membrane [I].Moreover, observations indicating an organ specificity of the adenine nucleotide translocator were made using Ouchterlony double diffusion and crossed immunoelectrophoretic methods [2, 31. However, the great hydrophobicity of this basic membrane protein necessitated several methodological modifications to ensure well-defined precipitation patterns on immunoelectrophoresis [4].For this reason and also to obtain further insight into the structure and function of the adenine nucleotide translocator (for review see [5,6]), we have elaborated an indirect solid phase radioimmunoassay in combination with immunoabsorption studies. These systems have the advantage that antigens and primary antibodies remain unmodified. Thus possible artefacts due to conformational changes...

show abstract

Section: Discussionmentioning

confidence: 99%

Immunochemical characterization of the adenine nucleotide translocator

Schultheiß

Klingenberg

1984

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…This was concluded from different apparent molecular mass [26,27], N-terminal amino acid sequence [25,28] and immunological reactivity [29] between corresponding nuclear encoded subunits of the liver and heart enzyme from the same animal.…”

mentioning

confidence: 99%

Isolation and properties of cytochrome c oxidase from rat liver and quantification of immunological differences between isozymes from various rat tissues with subunit-specific antisera

1985

View full text Add to dashboard Cite

Cytochrome c oxidase was isolated from rat liver either by affinity chromatography on cytochrome-cSepharose 4B or by chromatography on DEAE-Sepharose. Dodecyl sulfate gel electrophoresis of both preparations showed the same subunit pattern consisting of 13 different polypeptides. Kinetic analysis of the two preparations gave a higher V,,, for the enzyme isolated by chromatography on DEAE-Sephacel.Specific antisera were raised in rabbits against nine of the ten nuclear endoded subunits. A monospecific reaction of each antiserum with its corresponding subunit was obtained by Western blot analysis, thus excluding artificial bands in the gel electrophoretic pattern of the isolated enzyme due to proteolysis, aggregation or conformational modification of subunits.With an antiserum against rat liver holocytochrome c oxidase a different reactivity was found by Western blot analysis for subunits VIa and VIII between isolated cytochrome c oxidases from pig liver or kidney and heart or skeletal muscle.For a quantitative analysis of immunological differences a nitrocellulose enzyme-linked immunosorbent assay was developed. Monospecific antisera against 12 of the 13 subunits of rat liver cytochrome c oxidase were titrated with increasing amounts of total mitochondrial proteins from different rat tissues dissolved in dodecyl sulfate and dotted on nitrocellulose. The absorbance of a soluble dye developed by the second peroxidase-conjugated antibody was measured.From the data the following conclusions were obtained: (a) The mitochondrial encoded catalytic subunits

show abstract

“…These differences have been demonstrated to be due to different genes coding for these polypeptides in liver and heart. This was concluded from a very similar but slightly different N-terminal amino acid composition of the polypeptides from liver and heart [49].…”

Section: Discussionmentioning

confidence: 97%

Kinetic and Structural Differences between Cytochrome c Oxidases from Beef Liver and Heart

Merle

Kadenbach

1982

European Journal of Biochemistry

106

View full text Add to dashboard Cite

The cytochrome content of beef liver mitochondria differs from that of beef heart mitochondria by an eight‐ fold lower cytochrome aa3 and a twofold lower cytochrome b and c + c1 content. The kinetic properties of cytochrome c oxidases from beef liver and heart were measured with intact cytochrome c‐depleted membranes, deoxycholate‐dissolved membranes, and with the isolated enzymes at various cytochrome c concentrations with an oxygen electrode. Under all conditions a higher V was found for the liver enzyme, both for the low‐affinity and for the high‐affinity binding site for cytochrome c. Differences were also found for the Km of the two enzymes. Isolated beef heart mitochondria contained about twice as much cardiolipin than beef liver mitochondria. The isolated enzymes contained one mole cardiolipin per mole of the heart enzyme, but 2 moles cardiolipin per mole of the liver enzyme. By application of a high performance sodium dodecylsulfate gel electrophoretic system the two isolated enzymes could be separated into 13 different protein components, three of which (polypeptides VIa, VIIa and VIII) were found to differ in their apparent molecular weights. The functional meaning of cytochrome c oxidase iso‐ enzymes in liver and heart is discussed.

show abstract

Tissue‐specific genes code for polypeptide VIa of bovine liver and heart cytochrome c oxidase

Cited by 96 publications

References 10 publications

Immunochemical characterization of the adenine nucleotide translocator

Immunochemical characterization of the adenine nucleotide translocator

Isolation and properties of cytochrome c oxidase from rat liver and quantification of immunological differences between isozymes from various rat tissues with subunit-specific antisera

Kinetic and Structural Differences between Cytochrome c Oxidases from Beef Liver and Heart

Contact Info

Product

Resources

About