Biosynthesis of the major zona pellucida glycoprotein secreted by oocytes during mammalian oogenesis.
An antiserum directed specifically against ZP2, the major glycoprotein of the mouse egg's extracellular coat (zona pellucida), has been used to immunoprecipitate intracellular precursors of ZP2 that were synthesized by growing mouse oocytes cultured in vitro. Pulse-chase experiments revealed that the immediate precursor of mature, 120 kilodalton (kd) ZP2 is a 91 kd species that unlike mature ZP2, is sensitive to digestion by endo-beta-N-acetylglucosaminidase H (Endo H) and is converted by the endoglycosidase into an 81 kd species. An 81 kd species is only found intracellularly when growing oocytes are cultured in the presence of tunicamycin. These results suggest that ZP2 is synthesized as an 81 kd polypeptide chain that is first "core"-glycosylated at asparagine residues with high-mannose-type oligosaccharides, giving rise to a 91 kd intermediate (Endo-H-sensitive), and then processed to complex-type oligosaccharides prior to secretion as mature, 120 kd ZP2 (Endo H-insensitive). Furthermore, electrophoretic analyses of mature ZP2, ZP2 precursor (91 kd) and Endo H-treated ZP2 precursor (81 kd) suggest that there are six N-linked oligosaccharides per molecule and that the extreme heterogeneity of mature ZP2 is a consequence of the oligosaccharides and not the polypeptide chain itself.
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification; D007106:Immune Sera; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D051379:Mice; D009865:Oocytes / Q000378:metabolism*; D009866:Oogenesis; D010053:Ovary / Q000166:cytology; D010063:Ovum / Q000378:metabolism*; D015044:Zona Pellucida / Q000378:metabolism*
D006023:Glycoproteins; D007106:Immune Sera; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase