Isolation and characterization of membranes from oleic acid-induced peroxisomes of Candida tropicalis. 1990

W M Nuttley, and A G Bodnar, and D Mangroo, and R A Rachubinski
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.

We report a methodology for the isolation of peroxisome membranes from the yeast Candida tropicalis pK233 grown on oleic acid, and the characterization of the polypeptide and lipid compositions of these membranes. Peroxisomes purified in either sucrose or Nycodenz gradients are treated with Tris-HCl (pH 8.5) and then with sodium carbonate (pH 11.5) to yield a final peroxisome membrane preparation (hereafter called 'peroxisome membranes'). Electron microscopy revealed peroxisome membranes that are approximately 8.1 nm thick, have a typical trilaminar appearance, and form either flattened sheets or whorled structures. Peroxisome membranes contain 3.1% and 2.2% of the total protein of sucrose- and Nycodenz-gradient-purified peroxisomes, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed three predominant polypeptide bands of 34 (PMP 34), 29 (PMP 29), and 24 (PMP 24) x 10(3) Mr in peroxisome membranes. Immunoblotting with an antiserum to PMP 24 showed that PMP 24 segregates with the peroxisome membrane fractions and is induced by growth of Candida tropicalis on oleic acid. Peroxisome membranes contain neutral lipids and phospholipids. The principal phospholipids are phosphatidyl choline and phosphatidyl ethanolamine. The phospholipid/protein ratio of peroxisome membranes is approximately 430 nmol mg-1.

UI MeSH Term Description Entries
D007425 Intracellular Membranes Thin structures that encapsulate subcellular structures or ORGANELLES in EUKARYOTIC CELLS. They include a variety of membranes associated with the CELL NUCLEUS; the MITOCHONDRIA; the GOLGI APPARATUS; the ENDOPLASMIC RETICULUM; LYSOSOMES; PLASTIDS; and VACUOLES. Membranes, Intracellular,Intracellular Membrane,Membrane, Intracellular
D008563 Membrane Lipids Lipids, predominantly phospholipids, cholesterol and small amounts of glycolipids found in membranes including cellular and intracellular membranes. These lipids may be arranged in bilayers in the membranes with integral proteins between the layers and peripheral proteins attached to the outside. Membrane lipids are required for active transport, several enzymatic activities and membrane formation. Cell Membrane Lipid,Cell Membrane Lipids,Membrane Lipid,Lipid, Cell Membrane,Lipid, Membrane,Lipids, Cell Membrane,Lipids, Membrane,Membrane Lipid, Cell,Membrane Lipids, Cell
D008830 Microbodies Electron-dense cytoplasmic particles bounded by a single membrane, such as PEROXISOMES; GLYOXYSOMES; and glycosomes. Glycosomes,Glycosome,Microbody
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D009829 Oleic Acids A group of fatty acids that contain 18 carbon atoms and a double bond at the omega 9 carbon. Octadecenoic Acids,Acids, Octadecenoic,Acids, Oleic
D002175 Candida A genus of yeast-like mitosporic Saccharomycetales fungi characterized by producing yeast cells, mycelia, pseudomycelia, and blastophores. It is commonly part of the normal flora of the skin, mouth, intestinal tract, and vagina, but can cause a variety of infections, including CANDIDIASIS; ONYCHOMYCOSIS; VULVOVAGINAL CANDIDIASIS; and CANDIDIASIS, ORAL (THRUSH). Candida guilliermondii var. nitratophila,Candida utilis,Cyberlindnera jadinii,Hansenula jadinii,Lindnera jadinii,Monilia,Pichia jadinii,Saccharomyces jadinii,Torula utilis,Torulopsis utilis,Monilias
D002855 Chromatography, Thin Layer Chromatography on thin layers of adsorbents rather than in columns. The adsorbent can be alumina, silica gel, silicates, charcoals, or cellulose. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Thin-Layer,Thin Layer Chromatography,Chromatographies, Thin Layer,Chromatographies, Thin-Layer,Thin Layer Chromatographies,Thin-Layer Chromatographies,Thin-Layer Chromatography
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture

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