Purification of peroxisomes from livers of normal and clofibrate-treated mice

Crane, Denis I.; Hemsley, Anne C.; Masters, C.J.

doi:10.1016/0003-2697(85)90250-7

Cited by 31 publications

(9 citation statements)

References 21 publications

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“…These results indicated that while the peroxisomal matrix proteins function in specific metabolic pathways which are induced by specific substrates, the peroxisomal membrane proteins are more general and function in a substrate-nonspecific manner. Although peroxisomal proteins, from several organisms have been analyzed (10,21,25,28,42), functions for none of these resident proteins have been specifically defined. The gene that encodes PMP70 from rat liver peroxisomes has been reported (30), and it appears to have some similarity to a diverse family of proteins involved in membrane transport.…”

Section: Discussionmentioning

confidence: 99%

Association of glyoxylate and beta-oxidation enzymes with peroxisomes of Saccharomyces cerevisiae

et al. 1990

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Although peroxisomes are difficult to identify in Saccharomyces cerevisiae under ordinary growth conditions, they proliferate when cells are cultured on oleic acid. We used this finding to study the protein composition of these organelies in detail. Peroxisomes from oleic acid-grown cells were purified on a discontinuous sucrose gradient; they migrated to the 46 to 50% (wt/wt) sucrose interface. The peroxisomal fraction was identified morphologically and by the presence of all of the enzymes of the peroxisomal beta-oxidation pathway. These organelles also contained a significant but minor fraction of two enzymes of the glyoxylate pathway, malate synthase and malate dehydrogenase-2. The localization of malate synthase in peroxisomes was confirmed by immunoelectron microscopy. It is postulated that glyoxylate pathway enzymes are readily and preferentially released from peroxisomes upon cell lysis, accounting for their incomplete recovery from isolated organelles. Small uninduced peroxisomes from glycerol-grown cultures were detected on sucrose gradients by marker enzymes. Under these conditions, catalase, acyl-coenzyme A oxidase, and malate synthase cofractionated at equilibrium close to the mitochondrial peak, indicating smaller, less dense organelles than those from cells grown on oleic acid. Peroxisomal membranes from oleate cultures were purified by buoyant density centrifugation. Three abundant proteins of 24, 31, and 32 kilodaltons were observed.

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Section: Discussionmentioning

confidence: 99%

Association of glyoxylate and beta-oxidation enzymes with peroxisomes of Saccharomyces cerevisiae

et al. 1990

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“…Catalase activity in mouse liver fractions was assayed by a kinetic spectrophotometric procedure (6). The identities and levels of VLCFA in a total lipid extract from tissue samples were determined as previously described (33).…”

Section: S]methionine-labeled Protein From the Pex13mentioning

confidence: 99%

Pex13 Inactivation in the Mouse Disrupts Peroxisome Biogenesis and Leads to a Zellweger Syndrome Phenotype

Maxwell

Bjorkman

Nguyen

et al. 2003

Molecular and Cellular Biology

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107

111

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Zellweger syndrome is the archetypical peroxisome biogenesis disorder and is characterized by defective import of proteins into the peroxisome, leading to peroxisomal metabolic dysfunction and widespread tissue pathology. In humans, mutations in the PEX13 gene, which encodes a peroxisomal membrane protein necessary for peroxisomal protein import, can lead to a Zellweger phenotype. To develop mouse models for this disorder, we have generated a targeted mouse with a loxP-modified Pex13 gene to enable conditional Cre recombinasemediated inactivation of Pex13. In the studies reported here, we crossed these mice with transgenic mice that express Cre recombinase in all cells to generate progeny with ubiquitous disruption of Pex13. The mutant pups exhibited many of the clinical features of Zellweger syndrome patients, including intrauterine growth retardation, severe hypotonia, failure to feed, and neonatal death. These animals lacked morphologically intact peroxisomes and showed deficient import of matrix proteins containing either type 1 or type 2 targeting signals. Biochemical analyses of tissue and cultured skin fibroblasts from these animals indicated severe impairment of peroxisomal fatty acid oxidation and plasmalogen synthesis. The brains of these animals showed disordered lamination in the cerebral cortex, consistent with a neuronal migration defect. Thus, Pex13؊/؊ mice reproduce many of the features of Zellweger syndrome and PEX13 deficiency in humans.

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“…Peroxisomal membrane proteins with subunit molecular masses of 68kDa and 70kDa were isolated from peroxisomes purified from mouse liver [7]. Antisera against these proteins have been previously characterised and shown to be monospecific for these membrane proteins [6].…”

Section: Antiseramentioning

confidence: 99%

Ultrastructure and immunocytochemistry of hepatic peroxisomes in rhizomelic chondrodysplasia punctata

et al. 1992

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Peroxisomes were studied in the liver of two rhizomelic chondrodysplasia punctata patients using electron microscopy and catalase cytochemistry. Immunoelectron microscopy was carried out on the liver of one of these patients using antibodies to catalase, acyl-CoA oxidase, bifunctional protein, 3-ketoacyl-CoA thiolase and a 68 kDa peroxisomal membrane protein, in conjunction with protein-A colloidal gold. Moderately to markedly enlarged, flocculent peroxisomes were found in both patients. In one patient they were very heterogeneous with regard to the number per hepatocyte. The peroxisomes had very low levels of catalase as indicated by cytochemistry and immunocytochemistry. The three beta-oxidation enzymes were localised normally within the peroxisomes. The 68 kDa membrane protein was localised to the peroxisomal membranes. Some extra membrane loops were also identified using this antibody.

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Purification of peroxisomes from livers of normal and clofibrate-treated mice

Cited by 31 publications

References 21 publications

Association of glyoxylate and beta-oxidation enzymes with peroxisomes of Saccharomyces cerevisiae

Association of glyoxylate and beta-oxidation enzymes with peroxisomes of Saccharomyces cerevisiae

Pex13 Inactivation in the Mouse Disrupts Peroxisome Biogenesis and Leads to a Zellweger Syndrome Phenotype

Ultrastructure and immunocytochemistry of hepatic peroxisomes in rhizomelic chondrodysplasia punctata

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