Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
The N-ethylmaleimide-sensitive protein thiol groups necessary for sea-urchin egg cortical-granule exocytosis are highly exposed to the medium and are required for triggering by Ca2+.
Whalley T
,
Sokoloff A
.
???displayArticle.abstract???
It is known that sea-urchin egg cortical-granule exocytosis is inhibited by agents such as N-ethylmaleimide (NEM) which modify thiol groups. The fusion-related proteins modified by these agents have yet to be identified, nor is there information regarding the topography of these thiol groups. Furthermore, the step in cortical-granule exocytosis at which these thiol groups participate is unknown. In this study we have investigated the topological properties of, and the temporal requirement for the function of, the fusion-related thiol groups by treating the isolated exocytotic apparatus with high-molecular-mass dextrans and BSA carrying thiol-reactive 3-(2-pyridyldithio)propionate groups. The dextran derivatives inhibited exocytosis. The BSA derivative was much less inhibitory. Inhibition was reversed by treatment with dithiothreitol. When NEM was added to the dextran-derivative-treated exocytotic apparatus, treatment with dithiothreitol completely reversed inhibition, indicating that the dextran derivatives inhibit by reacting at the NEM-sensitive sites. A pulse of Ca2+ applied in the presence of inhibitors did not trigger any fusion following the removal of the inhibitor by dithiothreitol. These data show that the thiol groups, the modification of which by NEM inhibits exocytosis, are exposed to the medium in terms of their accessibility to macromolecules. They also show that the fusion-related thiol groups are required during the Ca(2+)-dependent stage of exocytosis.
Anderson,
Oocyte differentiation in the sea urchin, Arbacia punctulata, with particular reference to the origin of cortical granules and their participation in the cortical reaction.
1968, Pubmed,
Echinobase
Anderson,
Oocyte differentiation in the sea urchin, Arbacia punctulata, with particular reference to the origin of cortical granules and their participation in the cortical reaction.
1968,
Pubmed
,
Echinobase
Baker,
Influence of ATP and calcium on the cortical reaction in sea urchin eggs.
1978,
Pubmed
,
Echinobase
Beckers,
Vesicular transport between the endoplasmic reticulum and the Golgi stack requires the NEM-sensitive fusion protein.
1989,
Pubmed
Burgoyne,
Regulated exocytosis.
1993,
Pubmed
Carlsson,
Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent.
1978,
Pubmed
Chandler,
High molecular weight polymers block cortical granule exocytosis in sea urchin eggs at the level of granule matrix disassembly.
1989,
Pubmed
,
Echinobase
Chernomordik,
Lysolipids reversibly inhibit Ca(2+)-, GTP- and pH-dependent fusion of biological membranes.
1993,
Pubmed
,
Echinobase
Crossley,
Guanosine 5'-thiotriphosphate may stimulate phosphoinositide messenger production in sea urchin eggs by a different route than the fertilizing sperm.
1991,
Pubmed
,
Echinobase
Diaz,
Vesicle fusion following receptor-mediated endocytosis requires a protein active in Golgi transport.
1989,
Pubmed
Frye,
Arachidonic acid release and catecholamine secretion from digitonin-treated chromaffin cells: effects of micromolar calcium, phorbol ester, and protein alkylating agents.
1985,
Pubmed
Goda,
Identification of a novel, N-ethylmaleimide-sensitive cytosolic factor required for vesicular transport from endosomes to the trans-Golgi network in vitro.
1991,
Pubmed
Goud,
Small GTP-binding proteins and their role in transport.
1991,
Pubmed
Haggerty,
Release of granule contents from sea urchin egg cortices. New assay procedures and inhibition by sulfhydryl-modifying reagents.
1983,
Pubmed
,
Echinobase
Jackson,
Mild proteolytic digestion restores exocytotic activity to N-ethylmaleimide-inactivated cell surface complex from sea urchin eggs.
1985,
Pubmed
,
Echinobase
Jackson,
N-ethylmaleimide-sensitive protein(s) involved in cortical exocytosis in the sea urchin egg: localization to both cortical vesicles and plasma membrane.
1990,
Pubmed
,
Echinobase
Kaye,
Arachidonic acid-induced LH release is ATP-independent and insensitive to N-ethyl maleimide.
1992,
Pubmed
Lindau,
Techniques and concepts in exocytosis: focus on mast cells.
1991,
Pubmed
Luby-Phelps,
Behavior of a fluorescent analogue of calmodulin in living 3T3 cells.
1985,
Pubmed
Manoharan,
Spin label probes of the environment of cysteine beta-93 in hemoglobin.
1990,
Pubmed
Monsigny,
Colorimetric determination of neutral sugars by a resorcinol sulfuric acid micromethod.
1988,
Pubmed
Moy,
Calcium-mediated release of glucanase activity from cortical granules of sea urchin eggs.
1983,
Pubmed
,
Echinobase
O'Connor,
Synaptic vesicle exocytosis: molecules and models.
1994,
Pubmed
Plattner,
Stimulus-secretion coupling in Paramecium cells.
1991,
Pubmed
Popov,
Synaptotagmin: a calcium-sensitive inhibitor of exocytosis?
1993,
Pubmed
Rothman,
Molecular dissection of the secretory pathway.
1992,
Pubmed
Sasaki,
Modulation of calcium sensitivity by a specific cortical protein during sea urchin egg cortical vesicle exocytosis.
1984,
Pubmed
,
Echinobase
Sasaki,
Cortical vesicle exocytosis in isolated cortices of sea urchin eggs: description of a turbidometric assay and its utilization in studying effects of different media on discharge.
1983,
Pubmed
,
Echinobase
Steinhardt,
Intracellular calcium release at fertilization in the sea urchin egg.
1977,
Pubmed
,
Echinobase
Steinhardt,
Calmodulin confers calcium sensitivity on secretory exocytosis.
1982,
Pubmed
,
Echinobase
Swann,
The part played by inositol trisphosphate and calcium in the propagation of the fertilization wave in sea urchin eggs.
1986,
Pubmed
,
Echinobase
Söllner,
A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion.
1993,
Pubmed
Südhof,
Membrane fusion machinery: insights from synaptic proteins.
1993,
Pubmed
Tagaya,
Domain structure of an N-ethylmaleimide-sensitive fusion protein involved in vesicular transport.
1993,
Pubmed
Turner,
A cholera toxin-sensitive G-protein stimulates exocytosis in sea urchin eggs.
1987,
Pubmed
,
Echinobase
Turner,
Regulation of cortical vesicle exocytosis in sea urchin eggs by inositol 1,4,5-trisphosphate and GTP-binding protein.
1986,
Pubmed
,
Echinobase
Vacquier,
The isolation of intact cortical granules from sea urchin eggs: calcium lons trigger granule discharge.
1975,
Pubmed
,
Echinobase
Vogel,
Lysophosphatidylcholine reversibly arrests exocytosis and viral fusion at a stage between triggering and membrane merger.
1993,
Pubmed
,
Echinobase
Vogel,
Proteins on exocytic vesicles mediate calcium-triggered fusion.
1992,
Pubmed
,
Echinobase
Vogel,
Calcium-triggered fusion of exocytotic granules requires proteins in only one membrane.
1992,
Pubmed
,
Echinobase
Whalley,
Phosphoprotein inhibition of calcium-stimulated exocytosis in sea urchin eggs.
1991,
Pubmed
,
Echinobase
Whalley,
Exocytosis reconstituted from the sea urchin egg is unaffected by calcium pretreatment of granules and plasma membrane.
1988,
Pubmed
,
Echinobase
Whitaker,
Calcium-dependent exocytosis in an in vitro secretory granule plasma membrane preparation from sea urchin eggs and the effects of some inhibitors of cytoskeletal function.
1983,
Pubmed
,
Echinobase
Whitaker,
How calcium may cause exocytosis in sea urchin eggs.
1987,
Pubmed
,
Echinobase
Whitaker,
Inhibition of secretory granule discharge during exocytosis in sea urchin eggs by polymer solutions.
1987,
Pubmed
,
Echinobase
Zimmerberg,
Irreversible swelling of secretory granules during exocytosis caused by calcium.
1985,
Pubmed
,
Echinobase
