Isolation and Properties of Intact Mitochondria From Spheroplasts of Yeast

Duell, Elizabeth A.; Inoue, Sakae; Utter, Merton F.

doi:10.1128/jb.88.6.1762-1773.1964

Cited by 216 publications

(42 citation statements)

References 14 publications

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“…However, the presence of significant activities in non-mitochondrial fractions, and the alterations in these activities with the gene dosage of CYC3 demonstrate that the observed differences between strains do not result from variations in the details of the cell fractionations from strain to strain. In addition, differences between CYC3+ and cyc3strains comparable with those shown in Table I have been observed using mechanical disruption of cells (Mason et al, 1973) rather than the lysis of protoplasts (Duell et al, 1964) (results not shown).…”

Section: Demonstration That Cyc3 Encodes Cytochrome C Heme Lyasesupporting

confidence: 60%

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Identification and sequence of the gene encoding cytochrome c heme lyase in the yeast Saccharomyces cerevisiae.

Dumont¹,

Ernst²,

Hampsey³

et al. 1987

The EMBO Journal

231

175

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Mitochondrial cytochrome c contains a heme group covalently attached through thioether linkages to two cysteinyl residues of the protein. We demonstrate here that the nuclear gene, CYC3, in the yeast Saccharomyces cerevisiae, encodes cytochrome c heme lyase (CCHL), the enzyme catalyzing the attachment of heme to apocytochrome c. Mitochondrial extracts from cyc3‐ mutants are deficient in CCHL activity compared with extracts from normal strains, whereas strains carrying multiple copies of the CYC3 gene exhibit high levels of the activity. The CYC3 gene was cloned by functional complementation of a cyc3‐ mutant using a previously isolated plasmid containing the gene PYK1, which is tightly linked to CYC3. An open reading frame encoding a protein of 269 amino acids was identified from the DNA sequence of a fragment encompassing the CYC3 gene, and the corresponding transcript shown to be approximately 0.9 kb in length. CCHL appears to be a single polypeptide chain which acts specifically on the two forms of cytochrome c, but not on cytochrome c1.

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Section: Demonstration That Cyc3 Encodes Cytochrome C Heme Lyasesupporting

confidence: 60%

“…Mitochondrial fractions were isolated essentially as described by Duell et al (1964). After two washes in distilled water, a portion of the cells was set aside for the determination of total cellular protein by the boiling Biuret method of Strickland (1951), standardized against bovine serum albumin (BSA).…”

Section: Isolation Of Yeast Cellular Fractionsmentioning

confidence: 99%

Identification and sequence of the gene encoding cytochrome c heme lyase in the yeast Saccharomyces cerevisiae.

Dumont¹,

Ernst²,

Hampsey³

et al. 1987

The EMBO Journal

231

175

View full text Add to dashboard Cite

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Section: Isolation Of Mitochondriamentioning

confidence: 99%

“…Purified isolated mitochondria were prepared using the method of Duell et al [15]. This method, involving removal of the yeast cell wall using Glusulase to yield spheroplasts, was developed to yield high-quality mitochondria irrespective of the stage of growth of the yeast [15]. Because isoamyl alcohol-induced filaments were a novel source of mitochondria, preliminary experiments were performed to establish the optimum type of enzyme, quantity of enzyme required and the incubation time.…”

Section: Isolation Of Mitochondriamentioning

confidence: 99%

Isoamyl alcohol-induced morphological change in involves increases in mitochondria and cell wall chitin content

Kern

Nunn

Pichová

et al. 2004

FEMS Yeast Research

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Isoamyl alcohol reduced growth and induced filament formation in Saccharomyces cerevisiae. Isoamyl alcohol-induced filamentation was accompanied by an almost threefold greater increase in the specific activity of succinate dehydrogenase than in untreated cells, which suggested that isoamyl alcohol treatment caused the cells to produce more mitochondria than in normal yeast form proliferation. This was supported by measuring the dry weight of purified, isolated mitochondria. Filaments have an increased chitin content which is distributed over the majority of their surface, and is not confined to bud scars and the chitin ring between mother and daughter cells as in yeast-form cells.

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“…Mitochondria The transformation of yeast cells into spheroplasts mas performed by modification of the method described by Duel1 et al [9]. 5 g of derepresscd wet cells were preincubated in a solution of 0.14 M cysteamine-HC1 (pH 6.5) with 10mM EDTA for 30min a t 35" and centrifuged 5 min a t 30,000 x g. The pellet was suspended in 24 ml of 0.66 M sorbitol, buffered to p€I 5.8 by 5 mM citrate and 5 mM potassium phosphate, and 4 ml of Sue #Helix pomatia (containing 100,000 U p-glucuronidase and 800,000 U sulfatase/ ml), purchased from "Industrie Biologique Fran-Fake'' (Genevilliers, fiance).…”

Section: Formation Of Spheroplasts and Isolation Ofmentioning

confidence: 99%

Isoenzymes of Malate Dehydrogenase in Saccharomyces cerevisiae

Atzpodien¹,

Gancedo²,

Duntze³

et al. 1968

European Journal of Biochemistry

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Extracts from yeast cells grown under different conditions and from mitochondria were subjected to agar gel electrophoresis. Three bands showing malate dehydrogenase activity were found in extracts from aerobic derepressed cells (c-MDH I , c-MDH 11, and m-MDH 111), whereas generally only one band was present in extracts from anaerobic repressed cells (c-MDH 1) and another one in extracts from mitochondria (m-MDH 111) of aerobic derepressed cells.The difFerent malate dehydrogenases showed dissimilar regulatory and physical properties, e. g. sensitivity to inactivation by heat, inhibition by oxaloacetate and by p-chloromercuribenzoate, activation by Pi, Michaelis constants etc. A separation of malate dehydrogenases by DEAE-cellulose chromatography was performed giving only two malate dehydrogenase forms. These two forms were subjected to agar gel electrophoresis and gave one band each, corresponding to the cytoplasmic c-MDH I and c-MDH 11. The possible metabolic significance of the three malate dehydrogenase isoenzymes is discussed.

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Isolation and Properties of Intact Mitochondria From Spheroplasts of Yeast

Cited by 216 publications

References 14 publications

Identification and sequence of the gene encoding cytochrome c heme lyase in the yeast Saccharomyces cerevisiae.

Identification and sequence of the gene encoding cytochrome c heme lyase in the yeast Saccharomyces cerevisiae.

Isoamyl alcohol-induced morphological change in involves increases in mitochondria and cell wall chitin content

Isoenzymes of Malate Dehydrogenase in Saccharomyces cerevisiae

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