Synthesis and intracellular transport of cytochrome oxidase subunit IV and ADP/ATP translocator protein in intact hepatoma cells

Hatalová, Irena; Kolarov, Jordan

doi:10.1016/0006-291x(83)91270-6

Cited by 18 publications

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“…Processing of abundant mitochondrial matrix proteins was reported in liver explants [Il,13,141 and rat hepatocytes [15, 161. No information is available, however, on processing of the less abundant inner mitochondrial membrane peptides in mammalian cells. Recently, we employed a rat hepatoma ascites cell to investigate subunit IV of cytochrome oxidase [17]. In the present study, we have extended this system to investigate the formation and processing of cytochrome c1 and the iron sulfur protein of the cytochrome h-c, complex.…”

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Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells

Kolarov

Nelson

1984

European Journal of Biochemistry

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The import and processing of cytochrome c1 and the iron sulfur protein of the cytochrome h-c, complex were studied in Zajdela hepatoma ascites cells. Both peptides were synthesized as larger percursor molecules which were approximately 2-3 kDa and 5-6kDa larger than the mature forms of apocytochrome c1 and apo-iron sulfur protein, respectively. Comparison of these precursors to those reported for functionally homologous peptides in yeast and Neurospora indicate significant size changes have occurred in mammals.Rhodamine 6G, a specific vital stain for mitochondria, is a potent inhibitor of precursor processing in isolated hepatoma cells. Both precursor to cytochrome c1 and precursor to FeS accumulate in the soluble and particulate fractions obtained by digitonin treatment of tumor cells treated with Rhodamine 6G. Appearance of the mature peptides was abolished. The precursors are unstable, however, and disappear from the cytosolic and membrane fractions during a 10min chase. Comparison of the effects of Rhodamine 6G and carbonylcyanide m-chlorophenylhydrazone on precursor processing shows that: (a) Rhodamine 6G is a more effective inhibitor of processing, (b) it has less of an inhibitory effect on cellular protein synthesis, and (c) it inhibits processing under conditions in which it appears to have little influence on coupled respiration in whole cells. The data suggest that the most likely mode of action of Rhodamine 6G is on the matrix processing step.

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Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells

Kolarov

Nelson

1984

European Journal of Biochemistry

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“…In contrast, fewer inner membrane components which exhibit transmembrane activity have been studied. Three of these, the adenine nucleotide translocase (O'Malley et al, 1982;Hatalova & Kolarov, 1983), thermogenin (Freeman et al, 1983) and the phosphate-hydroxyl antiporter (G. M. Gibb & J. G. Lindsay, unpublished observations) are synthesized without a cleavable signal sequence. Whether the lack of such a signal sequence in these proteins is related to their topographical arrangement remains to be determined.…”

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Biosynthesis and processing of the large and small subunits of succinate dehydrogenase in cultured mammalian cells

Clarkson

King

Lindsay

1987

Biochemical Journal

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Monospecific polyclonal antisera have been raised to purified bovine heart succinate dehydrogenase and to the individual large and small subunits of this enzyme. These antisera exhibit cross-reactivity with the corresponding polypeptides in rat liver (BRL), pig kidney (PK-15) and bovine kidney (NBL-1) cell lines, and were employed to investigate some of the events involved in the biogenesis of succinate dehydrogenase in the PK-15 cell line. Newly-synthesized forms of the large and small subunits of succinate dehydrogenase were detected in cultured PK-15 and BRL cells labelled with [35S]methionine in the presence of uncouplers of oxidative phosphorylation. In PK-15 cells, the precursor forms of the large and small subunits exhibit Mr values approx. 1000-2000 and 4000-5000 greater than those of the corresponding mature forms. When the uncoupler is removed in pulse-chase experiments, complete conversion of the precursors to the mature forms occurs within 45 min. Studies on the kinetics of processing and stability of the large subunit precursor revealed that reversal of precursor accumulation is rapid, with processing occurring with a half-time of 5-7.5 min, and that the accumulated precursor exhibits long-term stability when PK-15 cells are maintained in the presence of 2,4-dinitrophenol.

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Characterization of two different genes (cDNA) for cytochrome c oxidase subunit VIa from heart and liver of the rat.

et al. 1988

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By antibody screening of a rat liver and a rat heart cDNA library in lambda gt11 two clones coding for the liver‐ and heart‐specific subunit VIa of rat cytochrome c oxidase were isolated. In the heart cDNA sequence a TAA stop codon was found in frame 18 bp 5′ upstream of the first methionine codon, thus excluding a leader sequence for this protein. The two cDNAs contain the full‐length coding region of two subunits. The amino acid sequences of the two subunits show only 50% homology, whereas 74% homology was found between rat heart and bovine heart subunit VIa. By Northern blot analysis it is shown that the gene for subunit VIa from heart is only expressed in heart and skeletal muscle, whereas that from liver is also expressed in kidney, brain, heart and weakly in muscle.

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Synthesis and intracellular transport of cytochrome oxidase subunit IV and ADP/ATP translocator protein in intact hepatoma cells

Cited by 18 publications

References 16 publications

Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells

Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells

Biosynthesis and processing of the large and small subunits of succinate dehydrogenase in cultured mammalian cells

Characterization of two different genes (cDNA) for cytochrome c oxidase subunit VIa from heart and liver of the rat.

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Synthesis and intracellular transport of cytochrome oxidase subunit IV and ADP/ATP translocator protein in intact hepatoma cells

Cited by 18 publications

References 16 publications

Import and processing of cytochrome b‐c1 complex subunits in isolated hepatoma ascites cells

Import and processing of cytochrome b‐c1 complex subunits in isolated hepatoma ascites cells

Biosynthesis and processing of the large and small subunits of succinate dehydrogenase in cultured mammalian cells

Characterization of two different genes (cDNA) for cytochrome c oxidase subunit VIa from heart and liver of the rat.

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Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells

Import and processing of cytochrome b‐c₁ complex subunits in isolated hepatoma ascites cells