Biosynthesis of the sperm receptor during oogenesis in the mouse.
During their growth phase, mouse oocytes synthesize and secrete three different glycoproteins, called ZP1, 2 and 3, that constitute the extracellular coat, or zona pellucida, of the oocyte. One of these glycoproteins, ZP3, exhibits properties expected for a sperm receptor. We have now used rabbit antisera that recognize ZP3 to immunoprecipitate [35S]methionine-labeled, intracellular precursors of this glycoprotein from growing oocytes cultured in vitro in the presence or absence of tunicamycin, a drug that prevents addition of N-linked oligosaccharides to nascent polypeptide chains. Electrophoretic analyses of these immunoprecipitates, as well as of immunoprecipitates digested with endo-beta-N-acetylglucosaminidase H (Endo H), indicate that ZP3 is synthesized as a 44,000 mol. wt. polypeptide chain to which either three or four high-mannose-type oligosaccharides are added, resulting in 53,000 and 56,000 mol. wt. ZP3 precursors, respectively. The latter species are converted to mature ZP3 (mol. wt. approximately 80,000) by processing of the high-mannose-type oligosaccharides (Endo H-sensitive) to complex-type oligosaccharides (Endo H-insensitive) prior to ZP3 secretion. The evidence presented reveals that the extreme heterogeneity of mature ZP3, with respect to both mol. wt. and isoelectric point, is partly a consequence of the N-linked oligosaccharides and not the polypeptide chain itself.
Salzmann
G S
GS
Department of Biological Chemistry, Harvard Medical School, Boston, MA 02115, USA.
Greve
J M
JM
Roller
R J
RJ
Wassarman
P M
PM
D000818:Animals; D004527:Egg Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D005260:Female; D006596:Hexosaminidases; D066298:In Vitro Techniques; D007526:Isoelectric Point; D008297:Male; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D051379:Mice; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D009866:Oogenesis / Q000502:physiology*; D011233:Precipitin Tests; D011498:Protein Precursors / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013084:Sperm-Ovum Interactions; D014415:Tunicamycin / Q000494:pharmacology; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011498:Protein Precursors; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; C000606519:Zp1 protein, mouse; C000606521:Zp3 protein, mouse; C035027:egg surface sperm receptor; D014415:Tunicamycin; D006596:Hexosaminidases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase