An Electron Transport Particle from Yeast: Purification and Properties

Mackler, Bruce; Collipp, P.J.; Duncan, Helen; Rao, N. Appaji; Huennekens, F.M.

doi:10.1016/s0021-9258(18)60258-3

Cited by 78 publications

(17 citation statements)

References 28 publications

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“…Succinate dehydrogenase (SDH) was assayed as INT reductase essentially as described by Nachlas et al (1960). NADH oxidase was assayed as described by Mackler et al (1962).…”

Section: Methodsmentioning

confidence: 99%

Formation of yeast mitochondria. VI. Mitochondriogenesis analyzed by blocks on mitochondrial translation and transcription

Mahler¹,

Perlman²

1971

Biochemistry

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The mitochondrial contribution to the biosynthesis of the respiratory chain in yeast has been analyzed by use of the inhibitors cycloheximide, chloramphenicol, and ethidium bromide. When care is taken to isolate primary from secondary effects, a minimal lesion can be localized in the region of cytochrome oxidase (enzymatic)/cytochrome aa3 (spectroscopic) ; the continued elaboration of these entities is abolished by both chloramphenicol and ethidium bromide added to either facultatively anaerobic petite-forming or to obligately * Contribution No.

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“…Succinate dehydrogenase (SDH) was assayed as INT reductase essentially as described by Nachlas et al (1960). NADH oxidase was assayed as described by Mackler et al (1962).…”

Section: Methodsmentioning

confidence: 99%

Formation of yeast mitochondria. VI. Mitochondriogenesis analyzed by blocks on mitochondrial translation and transcription

Mahler¹,

Perlman²

1971

Biochemistry

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“…The formation of H 2 O 2 was measured by recording the increase in absorbance at 500 nm. A mitochondrial marker enzyme, succinate-cytochrome C reductase, was assayed by the method of Mackler et al (1962). A lysosomal marker enzyme, acid phosphatase, was measured as described by Engström (1961).…”

Section: Enzyme Assaymentioning

confidence: 99%

Localization of Cytosolic NADP-dependent Isocitrate Dehydrogenase in the Peroxisomes of Rat Liver Cells

Yoshihara

Hamamoto

Munakata

et al. 2001

J Histochem Cytochem.

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S U M M A R YTwo types of NADP-dependent isocitrate dehydrogenases (ICDs) have been reported: mitochondrial (ICD1) and cytosolic (ICD2). The C-terminal amino acid sequence of ICD2 has a tripeptide peroxisome targeting signal 1 sequence (PTS1). After differential centrifugation of the postnuclear fraction of rat liver homogenate, approximately 75% of ICD activity was found in the cytosolic fraction. To elucidate the true localization of ICD2 in rat hepatocytes, we analyzed the distribution of ICD activity and immunoreactivity in fractions isolated by Nycodenz gradient centrifugation and immunocytochemical localization of ICD2 antigenic sites in the cells. On Nycodenz gradient centrifugation of the light mitochondrial fraction, ICD2 activity was distributed in the fractions in which activity of catalase, a peroxisomal marker, was also detected, but a low level of activity was also detected in the fractions containing activity for succinate cytochrome C reductase (a mitochondrial marker) and acid phosphatase (a lysosomal marker). We have purified ICD2 from rat liver homogenate and raised a specific antibody to the enzyme. On SDS-PAGE, a single band with a molecular mass of 47 kD was observed, and on immunoblotting analysis of rat liver homogenate a single signal was detected. Double staining of catalase and ICD2 in rat liver revealed co-localization of both enzymes in the same cytoplasmic granules. Immunoelectron microscopy revealed gold particles with antigenic sites of ICD2 present mainly in peroxisomes. The results clearly indicated that ICD2 is a peroxisomal enzyme in rat hepatocytes. ICD2 has been regarded as a cytosolic enzyme, probably because the enzyme easily leaks out of peroxisomes during homogenization.

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“…Compositional assays 5'-Nucleotidase (EC 3.1.3.5), Na+/K+-stimulated Mg2+ adenosine triphosphatase (EC 3.6.1.3) and NADH diaphorase (EC 1.6.99.3) were assayed as described by Avruch & Wallach (1971); glucose 6phosphatase (EC 3.1.3.9) was assayed by the method of Huibscher & West (1965) and succinate-cytochrome c reductase (EC 1.3.99.1) was assayed by the method of Mackler et al (1962). Protein was assayed by the method of Lowry et al (1951) with dry bovine serum albumin as standard.…”

Section: Resolution Ofmicrosomal Materialsmentioning

confidence: 99%

Isolation and characterization of membranes from normal and transformed tissue-culture cells

Graham

1972

Biochemical Journal

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Homogenates of baby-hamster kidney cells and rat embryo fibroblasts prepared by nitrogen cavitation contain a small population of slowly sedimenting mitochondria or mitochondrial fragments, which contaminate the microsomal fraction. This appears to limit the resolution of surface membrane and endoplasmic reticulum on magnesium-containing dextran gradients. The microsomal material and mitochondria can, however, be completely separated on a 10-60% (w/w) sucrose zonal gradient containing a 30% sucrose plateau. On magnesium-containing dextran gradients this mitochondria-free microsomal material can be resolved into at least two surface membrane fractions and at least two endoplasmic reticulum fractions. Comparison of polyoma virus-transformed and normal baby-hamster kidney cells reveals some interesting differences in their microsomal fractionation patterns and the characteristics of the Na(+)/K(+)-Mg(2+) adenosine triphosphatase of their surface membranes, in particular a tenfold lower K(m) in the virus-transformed cells. The fractionation patterns of normal and spontaneously transformed rat embryo fibroblasts are also briefly discussed, particularly in relation to the significance of the observation that both the surface membrane and endoplasmic reticulum from these cells can be subfractionated.

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An Electron Transport Particle from Yeast: Purification and Properties

Cited by 78 publications

References 28 publications

Formation of yeast mitochondria. VI. Mitochondriogenesis analyzed by blocks on mitochondrial translation and transcription

Formation of yeast mitochondria. VI. Mitochondriogenesis analyzed by blocks on mitochondrial translation and transcription

Localization of Cytosolic NADP-dependent Isocitrate Dehydrogenase in the Peroxisomes of Rat Liver Cells

Isolation and characterization of membranes from normal and transformed tissue-culture cells

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