Conversion of Ser to Thr residues at the sperm combining-site of mZP3 does not affect sperm receptor activity

Zev Williams, Eveline S. Litscher, Paul M. Wassarman

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Mammalian eggs are surrounded by a thick extracellular coat, the zona pellucida, that is composed of three glycoproteins, called ZP1-3. Sperm recognize and bind to O-linked oligosaccharides attached to Ser-332 and Ser-334 at the sperm combining-site of mouse ZP3 (mZP3). Mutation of either of these Ser residues to a small aliphatic amino acid results in the loss of sperm binding to mZP3 in vitro. Here, we converted both Ser-332 and Ser-334 to Thr residues by site-directed mutagenesis. Recombinant mutant glycoprotein made by stably transfected EC cells was purified and then assayed for its ability to inhibit binding of sperm to ovulated eggs in vitro. Results of these experiments suggest that Thr residues can replace the two evolutionarily conserved Ser residues as acceptors for essential O-linked oligosaccharides at the sperm combining-site of mZP3 without affecting the glycoprotein's sperm receptor activity.

Original languageEnglish (US)
Pages (from-to)813-818
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume301
Issue number4
DOIs
StatePublished - Feb 21 2003
Externally publishedYes

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Spermatozoa
Glycoproteins
Binding Sites
Oligosaccharides
Mutagenesis
Eggs
Zona Pellucida
Amino Acids
Site-Directed Mutagenesis
Fatty Acids
egg surface sperm receptor
Experiments
Mutation
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Conversion of Ser to Thr residues at the sperm combining-site of mZP3 does not affect sperm receptor activity. / Williams, Zev; Litscher, Eveline S.; Wassarman, Paul M.

In: Biochemical and Biophysical Research Communications, Vol. 301, No. 4, 21.02.2003, p. 813-818.

Research output: Contribution to journalArticle

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