Characterization of the zona pellucida glycoproteins from bovine ovarian and fertilized eggs

• Published in: Biochimica et biophysica acta Vol. 1201; no. 1; pp. 7 - 14
• Main Authors: Noguchi, Satoru, Yonezawa, Naoto, Katsumata, Toshiyuki, Hashizume, Ken-ichi, Kuwayama, Masashige, Hamano, Seizo, Watanabe, Shinya, Nakano, Minoru
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 28.09.1994; Elsevier
• Subjects: Amidohydrolases; amino acid composition; Amino Acid Sequence; amino acids; Amino Acids - analysis; analysis; Analytical, structural and metabolic biochemistry; Animals; beta-Galactosidase; Biological and medical sciences; Bovine egg; carbohydrate composition; Cattle; chemistry; Chromatography, Gel; composition; cows; Fertilization; fertilization (reproduction); Fundamental and applied biological sciences. Psychology; Glycoprotein; Glycoproteins; Glycoproteins - analysis; In vitro fertilization; Molecular Sequence Data; monosaccharides; Monosaccharides - analysis; ova; Ovum; Ovum - chemistry; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Proteins; Zona pellucida; Zona Pellucida - chemistry; In vitro fertilization; NeuGc; PBS; Glycoprotein; IEF; Zona pellucida; PVDF; Bovine egg; PMSF; ZP; NeuAc; EβG; TFMS; Ovary; Vertebrata; Mammalia; Bovine; Fertilization; N terminal-Sequence; Chemical composition; Glycoproteins; Artiodactyla; Two dimensional electrophoresis; Ungulata
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(94)90143-0

Bovine zone pellucida (ZP) glycorproteins from ovarian egg emerged as three bands with molecular mass of 78 kDa, 64 kDa and 21 kDa in SDS-PAGE under reducing conditions. Endo-β-galactosidase (EβG) digestion of the glycoproteins yielded five products with molecular mass of 76 kDa (EβG-76), 68 kDa (EβG-68), 63 kDa(EβG-63), 47 kDa (EβG-47) and 21 kDa (EβG-21) under the same conditions. The N-terminal amino acid sequences of EβG-76 and EβG-21 were identical. This fact together with the results of diagonal SDS-PAGE indicated that EβG-21 (N-terminal region) is linked to EβG-63 (C-terminal region) through disulfide bond to form EβG-76. Immunoblot analysis using anti-pig ZP protein antibodies revealed that bovine EβG-76, EβG-68 and EβG-47 correspond to pig PZP2, PZP3α and PZPEβ glycoproteins, respectively. The EβG-76 and EβG-68 components were shown to be specifically cleaved during fertilization.