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Identification of bovine zona pellucida glycoproteins
Author(s) -
Topper E.K.,
Kruijt L.,
Calvete J.,
Mann K.,
TöpferPetersen E.,
Woelders H.
Publication year - 1997
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199703)46:3<344::aid-mrd13>3.0.co;2-z
Subject(s) - zona pellucida , zona pellucida glycoprotein , biology , glycoprotein , gamete , peptide sequence , amino acid , microbiology and biotechnology , gel electrophoresis , sperm , oocyte , biochemistry , genetics , gene , embryo
Despite the economical importance of in vitro gamete technologies in cattle, only little is known about the molecular mechanisms of binding of spermatozoa to the zona pellucida (ZP) of the oocyte. The aim of the present work was to identify proteins from the bovine zona pellucida (bZP) and to investigate which bZP proteins play a role in sperm‐egg binding. High resolution 2‐dimensional polyacrylamide gel electrophoresis of bZP proteins under reducing conditions showed that the bovine ZP could be separated into 4 glycoprotein spots, provisionally named bZP1, bZP2, bZP3, and bZP4, with different molecular masses and isoelectrical points. The N‐terminal amino acid sequence of bZP1, bZP2, and bZP4 could be determined. The N‐terminal amino acid sequences of bZP1 and bZP4 were identical and were homologous to that of pZP4. Comparison of our data to that of Noguchi et al., 1994 (Biochim Biophys Acta 1201:7–14) revealed that bZP2 and bZP4 are fragments of bZP1. Immunoblot analysis showed that, respectively, anti‐porcine‐ZP3α and ‐ZP3β antibodies recognized 2 distinct regions of the bZP3 spot. Both antibodies inhibited sperm‐egg binding in the bovine. We conclude that the bovine ZP consists of 3 proteins that correspond by size, N‐terminal amino acid sequence, and antigenic determinants of pZP1, pZP3α, and pZP3β, respectively, that are encoded by the porcine ZPA, ZPB, and ZPC genes (Harris et al., 1994: J Seq Map 4:6331–393), respectively. Mol. Reprod. Dev. 46:344–350, 1997. © 1997 Wiley‐Liss, Inc.