Isolation of the zona pellucida and purification of its glycoprotein families from pig oocytes.
The isolation of the porcine zona pellucida, the glycoprotein envelope surrounding the mammalian oocyte, and the purification of its glycoprotein families is described. The zona pellucida was prepared from oocytes isolated from pig ovaries using a razor blade device and sieving procedures with Teflon or nylon screens. In 6-7 man h, the zona pellucida from 5 X 10(5) oocytes was obtained yielding 12 mg protein and 2.2 mg carbohydrate. The absorptivity of heat solubilized and filtered zona pellucida was A1%280 nm, 1 cm = 10.8. The four glycoprotein families of the zona pellucida were purified by two-dimensional polyacrylamide gel electrophoresis and electrophoretic elution. The electrophoretic purity of these families was greater than 90%. The protein and carbohydrate content and the amino acid and monosaccharide compositions of each of the glycoprotein families were determined.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods; D004527:Egg Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification*; D008562:Membrane Glycoproteins; D008970:Molecular Weight; D009865:Oocytes / Q000032:analysis*; D010063:Ovum / Q000032:analysis*; D011956:Receptors, Cell Surface; D013552:Swine; D015044:Zona Pellucida / Q000032:analysis*; D000072216:Zona Pellucida Glycoproteins
D000596:Amino Acids; D002241:Carbohydrates; D004527:Egg Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins