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ISOLATION OF POLAR GRANULES AND IDENTIFICATION OF POLAR GRANULE-SPECIFIC PROTEIN

DEVELOPMENTAL BIOLOGY 1978年第1期66卷 197-206页

作者： WARING, GL ALLIS, CD MAHOWALD, AP INDIANA UNIV DEPT ZOOL BLOOMINGTON IN 47401 USA

During early embryogenesis in Drosophila melanogaster the posterior polar plasm has the capacity to induce formation of primordial germ cells. Polar granules, organelles located exclusively in this polar plasm, were implicated in this determinative capacity. Using cell populations enriched for pole cells as starting material, a particulate subcellular fraction was obtained, that on EM analysis consisted predominately of polar granules. The chemical nature of the polar granule was defined by establishing a strict correlation between the morphological entity and the presence of specific chemical components. A basic protein with a MW on SDS[sodium dodecyl sulfate]-polyacrylamide gels of 95,000 daltons that was unique to embryonic cell populations containing pole cells was identified. This protein was enriched specifically in particulate subcellular fractions containing polar granules and was the only major protein species present in preparations in which polar granules were the major morphological constituent. Based on these data, polar granules appeared to be composed primarily of 1 major basic protein species.

关键词：细胞质颗粒/分析黑腹果蝇/胚胎学电泳聚丙烯酰氨凝胶胚胎非哺乳动物/细胞学显微镜检查电子分子量类器官/分析蛋白质类/分析亚细胞部分/分析动物

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SOME PROPERTIES OF NEUROPHYSINS ISOLATED FROM BOVINE NEUROSECRETORY GRANULES

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1975年第FEB21期248卷 235-245页

作者： PLISKA, V MEYERGRASS, M SWISS FED INST TECHNOL INST MOLEC BIOL & BIOPHYS8049 ZURICHSWITZERLAND

Our experiments with the neurophysin-related proteins from bovine NSG have demonstrated that these species differ in several important respects from the materials conventionally prepared. (1) In structural terms, the NSG proteins are essentially identical with the conventional neurophysins in amino acid composition and closely similar in immunoreactivity; however, the presence of carbodydrate and lipid moieties in the NSG material, no matter how they are attached, constitutes a structural difference apparently sufficient to cause considerable changes in properties. (2) The affinities for the neurohypophyseal hormones of the NSG proteins are very much higher than those of the 'conventional' neurophysins and, moreover, the binding properties of the NSG material are much more stable with time under the conditions of the binding experiments. (3) The low binding capacities of the NSG materials, even when they are purified to apparent molecular homogeneity, indicate a functional heterogeneity perhaps related to supramolecular structure. (4) The conversion of the NSG proteins by acid or alkali treatment to materials resembling the 'conventional' neurophysins in their binding properties suggests that the latter may be isolation artifacts. Although we cannot as yet consistently explain the properties of our neurophysins from NSG, we offer the hypothesis that the low binding capacity, as also the Hill coefficient greater than 1 (cf. Reference 25) are indicative of molecular aggregation, perhaps mediated or facilitated by the nonprotein components. It is conceivable that such aggregation, proceeding in the more 'natural' environment of the NSG in a more precisely organized manner, might constitute the truly 'native,' fully functional state of the neurophysins. In this context it is of interest to record our preliminary observations, which suggest the presence of a protein of 10,000 mol. wt. in the NSG membrane fraction (SDS gel) as well as electron-microscopic indications30 of

关键词：氨基酸类/分析结合部位交叉反应细胞质颗粒/分析电泳聚丙烯酰氨凝胶等电聚焦赖氨酸后叶激素运载蛋白类/分离和提纯垂体后叶/分析蛋白质结合蛋白质构象溶解度血管升压素类/类似物和衍生物动物牛

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PURIFICATION AND FRACTIONATION OF POTATO AMYLOPLASTS

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ANALYTICAL BIOCHEMISTRY 1975年第1-2期65卷 11-18页

作者： DUFFUS, CM ROSIE, R UNIV EDINBURGH SCH AGRDEPT AGR BIOCHEMEDINBURGH EH9 3JFSCOTLAND

A method is described for the isolation and purification of amyloplasts from mature potato tubers. The low-viscosity, high-density materials Urografin and Ludox TM were used in density gradients to obtain an additional fractionation on the grounds of size. Two main populations were identified. The properties of these materials and the purity of the amyloplast fractions are discussed.

关键词：直链淀粉/分析细胞分级分离/方法离心法梯密度细胞质颗粒/分析细胞质颗粒/超微结构 DNA/分析植物/分析植物/超微结构 RNA/分析

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BIOCHEMICAL AND MORPHOLOGICAL STUDIES OF RAT SUBMANDIBULAR-GLAND .1. CENTRIFUGAL FRACTIONATION OF GRANULE-RICH FRACTION

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JOURNAL OF DENTAL RESEARCH 1975年第5期54卷 938-947页

作者： HANKS, CT CHAKRABARTI, SG UNIV MICHIGAN SCH DENTINST DENT RESDEPT ORAL PATHOLANN ARBORMI 48104 UNIV MICHIGAN SCH DENTINST DENT RESDEPT ORAL BIOLANN ARBORMI 48104

Submandibular glands from male rats were homogenized in 0.34 M sucrose and 0.5 mM ethylenediaminetetraacetic acid in 10 mM HEPES buffer at a pH of 7.4. The extract was centrifuged and filtered through nuclepore filters to prepare a granule-rich fraction. Electron dense zymogen granules constituted approximately 85% of the particles in this fraction which also contained about a third of the total alkaline esterase activity in the gland.

关键词：酸性磷酸酶/代谢碱类离心法细胞质颗粒/分析细胞质颗粒/酶学细胞质颗粒/超微结构酯酶类/代谢酯类显微镜检查电子肽水解酶类/代谢亚细胞部分/酶学颌下腺/酶学颌下腺/超微结构动物男(雄)性大鼠

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SOLUBLE ACIDIC LIPOPROTEINS IN NEUROSECRETORY GRANULES - RELATION TO NEUROPHYSINS

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1975年第FEB21期248卷 218-234页

作者： KOENIG, H MYLROIE, R NORTHWESTERN UNIV MED SCHDEPT NEUROLCHICAGOIL 60611 VET ADM RES HOSP NEUROL SERVCHICAGOIL 60611

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CHARACTERIZATION OF RAT MAST-CELL GRANULE PROTEINS

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ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 1976年第2期173卷 554-563页

作者： LAGUNOFF, D PRITZL, P UNIV WASHINGTON SCH MED DEPT PATHOL SEATTLE WA 98195 USA

Mast cell granules free of membranes were isolated by differential centrifugation of water-lysed cells. The granules were extracted sequentially with 0.5, 1.0, and 2.0 M KCl. The 0.5 M fraction contained 95% of the N-acetyl-.beta.-glucosaminidase activity;this enzyme probably accounts for no more than 1% of the total granule protein. The 1.0 M fraction contained more than 80% of the granule chymotrypsin-like activity;the chymotrypsin-like enzyme was calculated to represent at least 15% of total granule protein. Heparin was found largely in the 1.0 M extract and in the residue after 2.0 M extraction. The heparin in both fractions had a molecular weight by gel exclusion chromatography considerably in excess of commercial porcine heparin. Acrylamide-gel electrophoresis of granules dissolved in 1% sodium dodecyl sulfate and reduced with dithiothreitol demonstrated 4 major protein bands. The 1.0 M fraction contained the most prominent, rapidly migrating band. The more slowly migrating, higher molecular weight bands appeared in grearer proportion in the 2.0 M and residue fractions. Autodigestion of the 1.0 M extract permitted purification of the mast cell chymotrypsin-like enzyme to specific activities as high as that of crystallized bovine pancreatic .alpha.-chymotrypsin. The mast cell chymotrypsin-like enzyme purified in this way migrated on dodecyl sulfate-gel electrophoresis as a single major band with an estimated molecular weight of 29,000.

关键词：细胞质颗粒/分析细胞质颗粒/超微结构电泳聚丙烯酰氨凝胶肥大细胞/分析肥大细胞/超微结构显微镜检查电子蛋白质类/分析动物男(雄)性大鼠

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LACTOSE AND MAJOR MILK-PROTEINS ARE PRESENT IN SECRETORY VESICLE-RICH FRACTIONS FROM LACTATING MAMMARY-GLAND

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1978年第10期75卷 5020-5024页

作者： SASAKI, M EIGEL, WN KEENAN, TW United States

Preparations enriched in apparently intact secretory vesicles were isolated from homogenates of lactating rat and bovine mammary tissue by differential and density gradient centrifugation in isoosmotic media. Morphologically, these preparations consisted nearly entirely of vesicles of varying sizes, at least some of which contained casein micelles. Endoplasmic reticulum vesicles, Golgi apparatus cisterna and dictyosomes, mitochondria, peroxisomes, lysosomes and nuclei were not observed in secretory vesicle-rich fractions. Vesicle preparations were enriched in lactose relative to total membrane fractions from mammary gland. The galactosyltransferase of lactose synthase (UDPgalactose: D-glucose 4-.beta.-galactosyl-transferase, EC 2.4.1.22) was also present in secretory vesicle preparations. .alpha.s1- and .beta.-caseins, .alpha.-lactalbumin and .beta.-lactoglobulin, the major secretory proteins of differentiated mammary epithelial cells, were identified as constituents of vesicle-rich fractions from bovine mammary gland. The major carbohydrate and major proteins of milk are apparently compartmentalized into secretory vesicles and are secreted by exocytotic fusion of secretory vesicles with the apical plasma membrane.

关键词：细胞区室化细胞分级分离细胞质颗粒/分析上皮/分析泌乳乳糖/分析乳糖合酶/分析乳腺动物/分析乳蛋白质类/分析动物牛女(雌)性妊娠大鼠

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POLYPHOSPHATE GRANULES IN ENCYSTED ZOOSPORES OF ROZELLA-ALLOMYCIS

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ARCHIVES OF MICROBIOLOGY 1976年第2期108卷 145-148页

作者： WOOL, SH HELD, AA CUNY HERBERT H LEHMAN COLL DEPT BIOL SCI BRONX NY 10468 USA ROCKEFELLER UNIV NEW YORK NY 10021 USA CUNY HERBERT H LEHMAN COLL DEPT BIOL SCI BRONX NY 10468 USA

Methods of ultracytochemistry and of X-ray energy dispersive analysis were used to demonstrate that the .gamma.-like granules in encysted zoospores of the chytrid R. allomycis contain polyphosphate. The possibility that cysts contain 2 classes of polyphosphate granules which differ in structure, in function and in origin is discussed.

关键词：细胞质颗粒/分析细胞质颗粒/超微结构真菌/超微结构卵菌纲/分析卵菌纲/超微结构磷酸盐类/分析孢子真菌/超微结构

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ORIGIN AND COMPOSITION OF CABOT RINGS IN PERNICIOUS-ANEMIA

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AMERICAN JOURNAL OF CLINICAL PATHOLOGY 1975年第1期64卷 53-57页

作者： KASS, L UNIV MICHIGAN SIMPSON MEM INST DEPT INTERN MED ANN ARBOR MI 48104 USA

Cytochemical studies were performed on erythrocytes containing Cabot rings from the peripheral blood of two patients with severe untreated pernicious anemia. These studies demonstrated that the Cabot ring contained arginine-rich histone and non-hemoglobin iron. Structures that may represent precursors of Cabot rings were found in stippled late intermediate marrow megaloblasts. The Cabot ring may result in part from abnormalities in metabolism of both iron and arginine-rich histone that are known to occur in pernicious anemia.

关键词：贫血恶性/病理学精氨酸/分析骨髓/病理学细胞质颗粒/分析 DNA/分析红细胞/分析红细胞/病理学组蛋白类/分析铁/分析赖氨酸/分析巨幼红细胞/病理学人类

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The lipid globules of saccharomyces cerevisiae. A combined chemical and ultrastructural study using ultrathin frozen sections

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Journal of Ultrasructure Research 1975年第1期51卷 32-39页

作者： Bauer, Heinz Sigarlakie, Elsje Bracco, Umberto Research and Development Department Nestlé Products Technical Assistance Co. Ltd. 1800 Vevey Switzerland

In unstained ultrathin frozen sections of Saccharomyces cerevisiae all lipid globules show an extremely high contrast due to a strong affinity towards OsO4. Uranyl-acetate/lead citrate poststaining of the sections con... 详细信息

In unstained ultrathin frozen sections of Saccharomyces cerevisiae all lipid globules show an extremely high contrast due to a strong affinity towards OsO₄. Uranyl-acetate/lead citrate poststaining of the sections considerably decreases the contrast of the lipid droplets. Two different modes of decolourization are observed: Part of the lipid globules are decolourized at their periphery only whereas others are decolourized homogeneously over their entire cross section. The presence of two different populations of lipid globules is therefore postulated. This is supported by freeze-etching results where two kinds of morphologically different lipid droplets are observed. Chemical analyses on the total lipid fraction revealed data commonly obtained for yeast cells. Even numbered monounsaturated fatty acids account for 70% of the total lipid fraction. Several possible explanations for the peculiar behaviour of the lipid globules during poststaining are forwarded. © 1975 Academic Press, Inc.

关键词：色谱法薄层细胞质颗粒/分析细胞质颗粒/超微结构脂肪酸类/分析冷冻蚀刻冷冻切片甘油酯类/分析组织学技术脂类/分析显微镜检查电子锇磷脂类/分析酿酒酵母菌/分析酿酒酵母菌/超微结构甾醇类/分析空泡/超微结构

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