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Abstract
The cell surface complex (Detering et al., 1977, J. Cell Biol. 75, 899-914) of the sea urchin egg consists of two subcellular organelles: the plasma membrane, containing associated peripheral proteins and the vitelline layer, and the cortical vesicles. We have now developed a method of isolating the plasma membrane from this complex and have undertaken its biochemical characterization. Enzymatic assays of the cell surface complex revealed the presence of a plasma membrane marker enzyme, ouabain-sensitive Na+/K+ ATPase, as well as two cortical granule markers, proteoesterase and ovoperoxidase. After separation from the cortical vesicles and purification on a sucrose gradient, the purified plasma membranes are recovered as large sheets devoid of cortical vesicles. The purified plasma membranes are highly enriched in the Na+/K+ ATPase but contain only very low levels of the proteoesterase and ovoperoxidase. Ultrastructurally, the purified plasma membrane is characterized as large sheets containing a "fluffy" proteinaceous layer on the external surface, which probably represent peripheral proteins, including remnants of the vitelline layer. Extraction of these membranes with Kl removes these peripheral proteins and causes the membrane sheets to vesiculate. Polyacrylamide gel electrophoresis of the cell surface complex, plasma membranes, and Kl-extracted membranes indicates that the plasma membrane contains five to six major proteins species, as well as a large number of minor species, that are not extractable with Kl. The vitelline layer and other peripheral membrane components account for a large proportion of the membrane-associated protein and are represented by at least six to seven polypeptide components. The phospholipid composition of the Kl-extracted membranes is unique, being very rich in phosphatidylethanolamine and phosphatidylinositol. Cholesterol was found to be a major component of the plasma membrane. Before Kl extraction, the purified plasma membranes retain the same species-specific sperm binding property that is found in the intact egg. This observation indicates that the sperm receptor mechanisms remain functional in the isolated, cortical vesicle-free membrane preparation.
BARTLETT,
Phosphorus assay in column chromatography.
1959, Pubmed
BARTLETT,
Phosphorus assay in column chromatography.
1959,
Pubmed
Castañeda,
Adenyl cyclase in plasma membrane preparations of sea urchin eggs and its increase in activity after fertilization.
1968,
Pubmed
,
Echinobase
Decker,
Sperm binding and fertilization envelope formation in a cell surface complex isolated from sea urchin eggs.
1979,
Pubmed
,
Echinobase
Detering,
Isolation and characterization of plasma membrane-associated cortical granules from sea urchin eggs.
1977,
Pubmed
,
Echinobase
FOLCH,
A simple method for the isolation and purification of total lipides from animal tissues.
1957,
Pubmed
Glabe,
Isolation and characterization of the vitelline layer of sea urchin eggs.
1977,
Pubmed
,
Echinobase
Heller,
The vitelline envelope of eggs from the giant keyhole limpet Megathura crenulata. I. Chemical composition and structural studies.
1976,
Pubmed
Herzog,
A new sensitive colorimetric assay for peroxidase using 3,3'-diaminobenzidine as hydrogen donor.
1973,
Pubmed
Kinsey,
Studies on the specificity of sperm binding in echinoderm fertilization.
1980,
Pubmed
,
Echinobase
LOWRY,
Protein measurement with the Folin phenol reagent.
1951,
Pubmed
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
MARSH,
The estimation of inorganic phosphate in the presence of adenosine triphosphate.
1959,
Pubmed
McBlaine,
Sea urchin egg hyaline layer: evidence for the localization of hyalin on the unfertilized egg surface.
1980,
Pubmed
,
Echinobase
Michell,
The possible involvement of phosphatidylinositol breakdown in the mechanism of stimulus-response coupling at receptors which control cell-surface calcium gates.
1977,
Pubmed
Renkonen,
The lipids of the plasma membranes and endoplasmic reticulum from cultured baby hamster kidney cells (BHK21).
1972,
Pubmed
Schmell,
Phospholipid metabolism in the eggs and embryos of the sea urchin Arbacia punctulata.
1974,
Pubmed
,
Echinobase
Schuel,
Sulfated acid mucopolysaccharides in the cortical granules of eggs. Effects of quaternary ammonium salts on fertilization.
1974,
Pubmed
,
Echinobase
Shapiro,
Limited proteolysis of some egg surface components is an early event following fertilization of the sea urchin, Strongylocentrotus purpuratus.
1975,
Pubmed
,
Echinobase
Turner,
Removal of lipid from intact erythrocytes and ghosts by aqueous solutions and its relevance to membrane structure.
1974,
Pubmed
Udenfriend,
Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range.
1972,
Pubmed
Veron,
Sequential biochemical and morphological events during assembly of the fertilization membrane of the sea urchin.
1977,
Pubmed
,
Echinobase
Wolf,
Isolation, physicochemical properties, and the macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs.
1976,
Pubmed
