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A method has been developed to isolate cortical granules (CG) free in suspension. It involves the mechanical disruption of the CG from CG lawns (CGL; Dev. Biol. 43:62-74, 1975) and concentration of the CG by low speed centrifugation. The isolated CG are intact and are a relatively pure population as judged by electron microscopy. Granule integrity is confirmed by the fact that isolated intact CG are radioiodinated to only 0.05% of the specific activity of hypotonically lysed CG. Purity of the CG preparation is assessed by the enrichment (four- to sevenfold) of CG marker enzymes and the absence or low activity of plasma membrane, mitochondrial, cytoplasmic, and yolk platelet marker enzyme activities. CG isolated from 125I-surface-labeled eggs have a very low specific radioactivity, demonstrating that CG contamination by the plasma membrane-vitelline layer (PM-VL) is minimal. CG yield is approximately 1% of the starting egg protein. The CG isolation method is simple and rapid, 4 mg of CG protein being obtained in 1 h. Isolated CG and PM-VL display distinct electrophoretic patterns on SDS gels. Actin is localized to the PM-VL, and all bands present in the CGL are accounted for in the CG and PM-VL. Calmodulin is associated with the CGL, CG, and PM-VL fractions, but is not specifically enriched in these fractions as compared with whole egg homogenates. This method of isolating intact CG from unfertilized sea urchin eggs may be useful for exploring the mechanism of Ca2+-mediated CG exocytosis.
Apps,
An adenosine triphosphatase isolated from chromaffin-granulate membranes is closely similar to F1-adenosine triphosphatase of mitochondria.
1979, Pubmed
Apps,
An adenosine triphosphatase isolated from chromaffin-granulate membranes is closely similar to F1-adenosine triphosphatase of mitochondria.
1979,
Pubmed Baker,
Influence of ATP and calcium on the cortical reaction in sea urchin eggs.
1978,
Pubmed
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Echinobase Begg,
pH regulates the polymerization of actin in the sea urchin egg cortex.
1979,
Pubmed
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Echinobase Bryan,
On the reconstitution of the crystalline components of the sea urchin fertilization membrane.
1970,
Pubmed
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Echinobase Burgess,
Characterization of calmodulin and calmodulin isotypes from sea urchin gametes.
1982,
Pubmed
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Echinobase Carroll,
Isolation and biological activity of the proteases released by sea urchin eggs following fertilization.
1975,
Pubmed
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Echinobase Castañeda,
Adenyl cyclase in plasma membrane preparations of sea urchin eggs and its increase in activity after fertilization.
1968,
Pubmed
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Echinobase Castle,
Studies of isolated secretion granules of the rabbit parotid gland: ion-induced changes in membrane integrity.
1981,
Pubmed Chambers,
The activation of sea urchin eggs by the divalent ionophores A23187 and X-537A.
1974,
Pubmed
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Echinobase Chandler,
Membrane fusion during secretion: cortical granule exocytosis in sex urchin eggs as studied by quick-freezing and freeze-fracture.
1979,
Pubmed
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Echinobase Charbonneau,
Purification of plant calmodulin by fluphenazine-Sepharose affinity chromatography.
1979,
Pubmed Cheung,
Purification and characterization of cyclic 3',5'-nucleotide phosphodiesterase from bovine brain.
1974,
Pubmed COOPERSTEIN,
A microspectrophotometric method for the determination of cytochrome oxidase.
1951,
Pubmed Creutz,
Self-association of synexin in the presence of calcium. Correlation with synexin-induced membrane fusion and examination of the structure of synexin aggregates.
1979,
Pubmed Creutz,
Secretory vesicle - cytosol interactions in exocytosis: isolation by Ca2+-dependent affinity chromatography of proteins that bind to the chromaffin granule membrane.
1981,
Pubmed Creutz,
A biophysical model of the chromaffin granule. Accurate description of the kinetics of ATP and Cl- dependent granule lysis.
1980,
Pubmed Creutz,
Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules.
1978,
Pubmed Decker,
Sperm binding and fertilization envelope formation in a cell surface complex isolated from sea urchin eggs.
1979,
Pubmed
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Echinobase DE DUVE,
Distribution of enzymes between subcellular fractions in animal tissues.
1962,
Pubmed Detering,
Isolation and characterization of plasma membrane-associated cortical granules from sea urchin eggs.
1977,
Pubmed
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Echinobase Doré,
Acid phosphatase analysis in sea urchin eggs and blastulae.
1967,
Pubmed
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Echinobase Epel,
Calmodulin activates NAD kinase of sea urchin eggs: an early event of fertilization.
1981,
Pubmed
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Echinobase Epel,
Beta-1,3-glucanase of sea urchin eggs: release from particles at fertilization.
1969,
Pubmed
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Echinobase Fodor,
Isolation of a protease from sea urchin eggs before and after fertilization.
1975,
Pubmed
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Echinobase Foerder,
Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with tyrosine crosslinks.
1977,
Pubmed
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Echinobase Gilkey,
A free calcium wave traverses the activating egg of the medaka, Oryzias latipes.
1978,
Pubmed
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Echinobase Grab,
Presence of calmodulin in postsynaptic densities isolated from canine cerebral cortex.
1979,
Pubmed Hall,
Hardening of the sea urchin fertilization envelope by peroxidase-catalyzed phenolic coupling of tyrosines.
1978,
Pubmed
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Echinobase Head,
Calcium-binding modulator protein from the unfertilized egg of the sea urchin Arbacia punctulata.
1979,
Pubmed
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Echinobase Heuser,
Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release.
1979,
Pubmed Hoffman,
ATP-stimulated transmitter release and cyclic AMP synthesis in isolated chromaffin granules.
1976,
Pubmed Hubbard,
The enzymatic iodination of the red cell membrane.
1972,
Pubmed Hubbard,
Externally disposed plasma membrane proteins. I. Enzymatic iodination of mouse L cells.
1975,
Pubmed Johnson,
Proton: substrate stoichiometries during active transport of biogenic amines in chromaffin ghosts.
1981,
Pubmed Kane,
Preparation and purification of polymerized actin from sea urchin egg extracts.
1975,
Pubmed
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Echinobase Katsura,
Peroxidatic activity of catalase in the cortical granules of sea urchin eggs.
1974,
Pubmed
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Echinobase Kinsey,
Isolation and partial characterization of the plasma membrane of the sea urchin egg.
1980,
Pubmed
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Echinobase KRAHL,
Glucose-6-phosphate and 6-phosphogluconate dehydrogenases from eggs of the sea urchin, Arbacia punctulata.
1955,
Pubmed
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Echinobase Lawson,
Molecular events during membrane fusion. A study of exocytosis in rat peritoneal mast cells.
1977,
Pubmed Lazarides,
Actin is the naturally occurring inhibitor of deoxyribonuclease I.
1974,
Pubmed Lin,
Purification and characterization of a protein activator of cyclic nucleotide phosphodiesterase from bovine brain.
1974,
Pubmed Linden,
Interactions of calmodulin with coated vesicles from brain.
1981,
Pubmed Lopo,
Radioiodination and characterization of the plasma membrane of sea urchin sperm.
1980,
Pubmed
,
Echinobase LOWRY,
Protein measurement with the Folin phenol reagent.
1951,
Pubmed Nishida,
Calcium sensitivity of sea urchin tubulin in in vitro assembly and the effects of calcium-dependent regulator (CDR) proteins isolated from sea urchin eggs and porcine brains.
1980,
Pubmed
,
Echinobase Njus,
Phosphorus-31 nuclear magnetic resonance studies of active proton translocation in chromaffin granules.
1978,
Pubmed Pazoles,
Evidence for stimulation of anion transport in ATP-evoked transmitter release from isolated secretory vesicles.
1978,
Pubmed Plattner,
Genetic dissection of the final exocytosis steps in Paramecium tetraurelia cells: cytochemical determination of Ca2+-ATPase activity over performed exocytosis sites.
1980,
Pubmed Pollard,
The chromaffin granule and possible mechanisms of exocytosis.
1979,
Pubmed Roffman,
Sensitive proteolytic enzyme assay using differential solubilities of radioactive substrates and products in biphasic systems.
1970,
Pubmed Satir,
Membrane fusion in a model system. Mucocyst secretion in Tetrahymena.
1973,
Pubmed Scharschmidt,
Isolation of a rat liver plasma membrane fraction of probable canalicular origin. Preparative technique, enzymatic profile, composition, and solute transport.
1981,
Pubmed Schatten,
The penetration of the spermatozoon through the sea urchin egg surface at fertilization. Observations from the outside on whole eggs and from the inside on isolated surfaces.
1976,
Pubmed
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Echinobase Schuel,
A trypsin-like proteinase localized in cortical granules isolated from unfertilized sea urchin eggs by zonal centrifugation. Role of the enzyme in fertilization.
1973,
Pubmed
,
Echinobase Schuel,
Separation of intact cortical granules from homogenate of unfertilized sea urchin eggs by zonal centrifugation.
1969,
Pubmed
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Echinobase Schuel,
Purification of cortical granules from unfertilized sea urchin egg homogenates by zonal centrifugation.
1972,
Pubmed
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Echinobase SHIBKO,
Acid phosphatase of the lysosomal and soluble fraction of rat liver.
1963,
Pubmed SMITH,
Spectrophotometric assay of cytochrome c oxidase.
1955,
Pubmed Spudich,
Biochemical and structural studies of actomyosin-like proteins from non-muscle cells. II. Purification, properties, and membrane association of actin from amoebae of Dictyostelium discoideum.
1974,
Pubmed Spudich,
Actin in triton-treated cortical preparations of unfertilized and fertilized sea urchin eggs.
1979,
Pubmed
,
Echinobase Steinhardt,
Intracellular calcium release at fertilization in the sea urchin egg.
1977,
Pubmed
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Echinobase Steinhardt,
Calmodulin confers calcium sensitivity on secretory exocytosis.
1982,
Pubmed
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Echinobase Steinhardt,
Is calcium ionophore a universal activator for unfertilised eggs?
1974,
Pubmed
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Echinobase Strewler,
Purification and characterization of phosphodiesterase activator from kidney. A lysosomal protease.
1979,
Pubmed Talbot,
The purification and characterization of an exo-beta (1 going to 3)-glucanohydrolase from sea urchin eggs.
1982,
Pubmed
,
Echinobase Tashima,
Removal of protein interference in the Fiske-Subbarow method by sodium dodecyl sulfate.
1975,
Pubmed Tucker,
The effect of proteolysis on the calmodulin activation of cyclic nucleotide phosphodiesterase.
1981,
Pubmed Vacquier,
The isolation of intact cortical granules from sea urchin eggs: calcium lons trigger granule discharge.
1975,
Pubmed
,
Echinobase Vacquier,
The cytolytic isolation of the cortex of the sea urchin egg.
1980,
Pubmed
,
Echinobase Vacquier,
Calcium activation of esteroproteolytic activity obtained from sea urchin egg cortical granules.
1975,
Pubmed
,
Echinobase Vacquier,
Dynamic changes of the egg cortex.
1981,
Pubmed Wu,
Two proteins from cowpea mosaic virus.
1971,
Pubmed Zucker,
Prevention of the cortical reaction in fertilized sea urchin eggs by injection of calcium-chelating ligands.
1978,
Pubmed
,
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