Phospholipid analysis on a micro scale.
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Biomedical subjects
Publications and source records attributed to H Kleinig.
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The cell wall of the marine chrysophycean alga Pleurochrysis scherfellii is composed of distinct wall fragments embedded in a gelatinous mass. The latter is a polysaccharide of pectic character which is rich in galactose and ribose. These wall fragments are identified as scales. They have been isolated and purified from the vegetative mother cell walls after zoospore formation. Their ultrastructure is described in an electron microscope study combining sectioning, freeze-etch, and negative staining techniques. The scales consist of a layer of concentrically arranged microfibrils (ribbons with cross-sections of 12 to 25 x 25 to 40 A) and underlying radial fibrils of similar dimensions. Such a network-plate is densely coated with particles which are assumed to be identical to the pectic component. The microfibrils are resistant to strong alkaline treatment and have been identified as cellulose by different methods, including sugar analysis after total hydrolysis, proton resonance spectroscopical examination (NMR spectroscopy) of the benzoylated product, and diverse histochemical tests. The formation and secretion of the scales can be followed along the maturing Golgi cisternae starting from a pronounced dilated "polymerization center" as a completely intracisternal process which ends in the exocytotic extrusion of the scales. The scales reveal the very same ultrastructure within the Golgi cisternae as they do in the cell wall. The present finding represents the first evidence on cellulose formation by the Golgi apparatus and is discussed in relation to a basic scheme for cellulose synthesis in plant cells in general.
Nuclear membranes were isolated from rat and pig liver by sonication of highly purified nuclear fractions and subsequent removal of adhering nucleoproteins in a high salt medium. The fractions were examined in the electron microscope by both negative staining and thin sectioning techniques and were found to consist of nuclear envelope fragments of widely varying sizes. Nuclear pore complex constituents still could frequently be recognized. The chemical composition of the nuclear membrane fractions was determined and compared with those of microsomal fractions prepared in parallel. For total nuclei as well as for nuclear membranes and microsomes, various enzyme activities were studied. The results indicate that a similarity exists between both fractions of cytomembranes, nuclear envelope, and endoplasmic reticulum, with respect to their RNA:protein ratio and their content of polar and nonpolar lipids. Both membranous fractions had many proteins in common including some membrane-bound enzymes. Activities in Mg-ATPase and the two examined cytochrome reductases were of the same order of magnitude. The content of cytochrome b(5) as well as of P-450 was markedly lower in the nuclear membranes. The nuclear membranes were found to have a higher buoyant density and to be richer in protein. The glucose-6-phosphatase and Na-K-ATPase activities in the nuclear membrane fraction were very low. In the gel electrophoresis, in addition to many common protein bands, some characteristic ones for either microsomal or nuclear membranous material were detected. Significant small amounts of DNA and RNA were found to remain closely associated with the nuclear envelope fragments. Our findings indicate that nuclear and endoplasmic reticulum membranes which are known to be in morphological continuity have, besides a far-reaching similarity, some characteristic differences.
The qualitative and quantitative lipid composition of nuclei and nuclear membranes from pig and rat liver were determined. These determinations were compared with the corresponding data obtained for microsomes from the same material after similar treatments. The results indicate that, at least, by far the major part of the nuclear lipids is located in the membranes of the nuclear envelope. The phospholipid pattern of the nuclear membranes and the endoplasmic reticulum (ER) membranes in general is widely identical in both species. As a striking difference in the lipid composition, however, a fourfold increase of esterified cholesterol in the nuclear membranes was found. In a quantitative approach the ratio of total surface area of the nuclear lipids to the total surface area of the nuclear envelope membranes was calculated as being 3.6, a value which fairly approximates the requirements of a bimolecular lipid leaflet model.
The Golgi apparatus of a marine chrysophycean alga Pleurochrysis scherffelii Pringsheim produces wall fragments (circular-to-ellipsoidal "scales") which are released to the periphery by an exocytotic process involving the fusion of cisternae and the plasma membrane. The cellulosic component of the scales is a complex network of fibrils (10 to 25 angstroms in diameter) that resist treatment with strong alkali. Untreated washed scales yield galactose, ribose, arabinose, and traces of glucose; alkali-purified scales yield much more glucose. The fibrillar scale constituent shows a positive iodine dichroism of the intact wall, a positive zinc chloride-iodine reaction, breakage sites characteristic of highly crystalline cellulose, and solubility in Schweizer's reagent.
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