Subcellular fractionation by differential and zonal centrifugation of aerobically grown glucose-de-repressed <i>Saccharomyces carlsbergensis</i>

Cartledge, Trevor G.; Lloyd, David

doi:10.1042/bj1260381

Cited by 41 publications

(36 citation statements)

References 29 publications

(27 reference statements)

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“…(3) APytchromec oxidoreductases7. 8%, cytochrome c oxidase and (10) (Cartledge & Lloyd, 1972a). Fraction 39 also contained small electron-dense vesicles which appeared to be identical with those of fraction 43 of cells adapted for 10min.…”

Section: Fractionation Of Homogenates Of Cells After 30min Respiratormentioning

confidence: 73%

“…After 3h adaptation these mitochondrial marker enzymes were almost entirely confined to a zone at p = 1.23 g/ml, which corresponds to the density of mitochondria in aerobically grown, glucose-de-repressed cells (Cartledge & Lloyd, 1972a).…”

Section: Discussionmentioning

confidence: 92%

“…132 hexylcarbodi-imide (Fig. 3X) (Cartledge & Lloyd, 1972a). Fraction 46 also contained mitochondria-like organelles (Plate ld).…”

Section: Fractionation Of Homogenates Of Cells After 10min Respiratormentioning

confidence: 95%

“…Preparation of sphaeroplasts and subsequent disruption and centrifugation procedures for the production ofwhole homogenates used in all cell-fractionation studies were identical with those described by Cartledge & Lloyd (1972a).…”

Section: Harvesting Of Cells Andpreparation Ofextractsmentioning

confidence: 99%

“…Fractionation by high-speed zonal centrifugation Zonal centrifugations were performed in a B XIV zonal rotor as detailed previously (Cartledge & Lloyd, 1972a (EC 3.6.1.4) activities were assayed as described previously (Cartledge & Lloyd, 1972a).…”

Section: Harvesting Of Cells Andpreparation Ofextractsmentioning

confidence: 99%

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Changes in enzyme activities and distributions during glucose de-repression and respiratory adaptation of anaerobically grown Saccharomyces carlsbergensis

Cartledge¹,

Lloyd²

1973

Biochemical Journal

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1. During anaerobic glucose de-repression the respiration rate of whole cells of Saccharomyces carlsbergensis remained constant and was insensitive to antimycin A but was inhibited by 30 % by KCN. Aeration of cells for 1 h led to increased respiration rate which was inhibited by 80% by antimycin A or KCN. 2. Homogenates were prepared from sphaeroplasts of anaerobically grown, glucose de-repressed cells and the distribution of marker enzymes was investigated after zonal centrifugation on sucrose gradients containing MgCl2. These homogenates contained no detectable cytochrome c oxidase or catalase activity. The complex density distributions of NADH-and NADPH-cytochrome c oxidoreductases and adenosine triphosphatase(s) [ATPase(s)] were very different from those of anaerobically grown, glucose-repressed cells. 3. The specific activity of total ATPase was lowered and sensitivity to oligomycin decreased from 58 to 7 % during de-repression. 4. Cytochrome c oxidase and catalase activities were detectable in homogenates of cells after 10min aeration. Zonal centrifugation indicated complex, broad sedimentable distributions of all enzyme activities assayed; the peaks of activity were at 1.27g/ml. 5. Centrifugation of homogenates of cells adapted for 30min and 3h indicated a shift of density of the major sedimentable peak from 1.25g/ml (30min) to 1.235g/ml (3h). After 30min adaptation a minor zone of oligomycinsensitive ATPase and 15 % of the total cytochrome c oxidase activities were detected at p = 1.12g/l; these particles together with those of higher density containing cytochrome c oxidase, ATPase and NADH-cytochrome c oxidoreductase activities were all sedimented at 105g-min. 6. Electron microscopy indicated that the mitochondria-like structures of anaerobically grown, glucose-de-repressed cells were similar to those of repressed cells. After 10min of respiratory adaptation highly organized mitochondria were evident which resembled the condensed forms of mitochondria of aerobically grown, glucose-derepressed cells. High-density zonal fractions of homogenates of cells after adaptation also contained numerous electron-dense vesicles 0.05-0.2,um in diameter. 7. The possibility that the 'promitochondria' of anaerobically grown cells may not be the direct structural precursors of fully functional mitochondria is discussed.

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Section: Fractionation Of Homogenates Of Cells After 30min Respiratormentioning

confidence: 73%

Section: Discussionmentioning

confidence: 92%

“…132 hexylcarbodi-imide (Fig. 3X) (Cartledge & Lloyd, 1972a). Fraction 46 also contained mitochondria-like organelles (Plate ld).…”

Section: Fractionation Of Homogenates Of Cells After 10min Respiratormentioning

confidence: 95%

Section: Harvesting Of Cells Andpreparation Ofextractsmentioning

confidence: 99%

Section: Harvesting Of Cells Andpreparation Ofextractsmentioning

confidence: 99%

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Changes in enzyme activities and distributions during glucose de-repression and respiratory adaptation of anaerobically grown Saccharomyces carlsbergensis

Cartledge¹,

Lloyd²

1973

Biochemical Journal

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Location of three key enzymes of gluconeogenesis in baker's yeast

Haarasilta

Taskinen

1977

Arch. Microbiol.

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The subcellular location of hexose diphosphatase, phosphoenolpyruvate carboxykinase and pyruvate carboxylase in baker's yeast (Saccharomyces cerevisiae) was investigated by density gradient centrifugation of spheroplast lysates obtained by osmotic shock treatment of spheroplasts and centrifugation for 10000 g x min. On the evidence obtained from zonal separations these three enzymes of gluconeogenesis are most probably located in the soluble cytosol.

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Transmembrane ferricyanide reduction by cells of the yeastSaccharomyces cerevisiae

Crane

Roberts

Linnane

et al. 1982

J Bioenerg Biomembr

102

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Both respiratory-competent and respiratory-deficient yeast cells reduce external ferricyanide. The reduction is stimulated by ethanol and inhibited by the alcohol dehydrogenase inhibitor, pyrazole. The reduction of ferricyanide is not inhibited by inhibitors of mitochondrial or microsomal ferricyanide reduction. Cells in exponential-phase growth show a much higher rate of ferricyanide reduction. The reduction of ferricyanide is accompanied by increased release of protons by the yeast cells. We propose that the ferricyanide reduction is carried out by a transmembrane NADH dehydrogenase.

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Subcellular fractionation by differential and zonal centrifugation of aerobically grown glucose-de-repressed Saccharomyces carlsbergensis

Cited by 41 publications

References 29 publications

Changes in enzyme activities and distributions during glucose de-repression and respiratory adaptation of anaerobically grown Saccharomyces carlsbergensis

Changes in enzyme activities and distributions during glucose de-repression and respiratory adaptation of anaerobically grown Saccharomyces carlsbergensis

Location of three key enzymes of gluconeogenesis in baker's yeast

Transmembrane ferricyanide reduction by cells of the yeastSaccharomyces cerevisiae

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