Subcellular fractionation by zonal centrifugation of glucose-repressed anaerobically grown <i>Saccharomyces carlsbergensis</i>

Cartledge, Trevor G.; Lloyd, David

doi:10.1042/bj1270693

Cited by 20 publications

(15 citation statements)

References 15 publications

(32 reference statements)

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“…Fraction 41 contained ellipsoid mitochondria-like structures 0.3-0.5,um in length (Plate la), which showed poorly developed inner membranes and were similar in appearance to structures found in anaerobically grown, glucose-repressed cells (Cartledge & Lloyd, 1972c).…”

Section: Resultsmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 91%

“…(Cartledge & Lloyd, 1972c) had increased from p 1.235 to p = 1.27g/ml. Comparison of electron micrographs of the mitochondrialike organelles found in the anaerobic glucoserepressed state (Cartledge & Lloyd, 1972c) with those presented here for the glucose-de-repressed organelles indicates that no marked morphological changes have accompanied this increased buoyant density. After only 10min of respiratory adaptation cytochrome c oxidase was present in functional electrontransport chains and was found in particles large enough to be completely sedimented in 0.25 M-sucrose after centrifugation for 10'g-min (Cartledge et al, 1972).…”

Section: Discussionmentioning

confidence: 91%

“…74S) was maintained and grown under strictly anaerobic conditions in a complex growth medium in the presence of 10% (w/v) glucose and lipid supplements (Tween 80 and ergosterol) as previously described (Cartledge & Lloyd, 1972c).…”

Section: Materials and Methods Maintenance And Growthmentioning

confidence: 99%

“…Cells were harvested and, for the determination of total enzyme activities, extracts were prepared by using a Hughes Press (Hughes, 1951) as described previously (Cartledge & Lloyd, 1972c). 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%

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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: Resultsmentioning

confidence: 67%

Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 91%

Section: Materials and Methods Maintenance And Growthmentioning

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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Spezielle Cytologic: Morphologie und Morphogenese von Pilzzellen

Girbardt¹

1973

Fortschritte Der Botanik

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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 zonal centrifugation of glucose-repressed anaerobically grown Saccharomyces carlsbergensis

Cited by 20 publications

References 15 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

Spezielle Cytologic: Morphologie und Morphogenese von Pilzzellen

Transmembrane ferricyanide reduction by cells of the yeastSaccharomyces cerevisiae

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