Tyrosine phosphorylation of a 95 kDa protein and induction of the acrosome reaction in human spermatozoa by recombinant human zona pellucida glycoprotein 3

I. A. Brewis, R. Clayton, C. E. Browes, M. Martin, C. L. R. Barratt, D. P. Hornby, H. D. M. Moore

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

Protein tyrosine phosphorylation and induction of the acrosome reaction (AR) in non-capacitated and capacitated human spermatozoa was investigated in response to recombinant human zona pellucida glycoprotein (rhZP3) produced by Chinese hamster ovary cells transfected with a plasmid containing human ZP3 cDNA. rhZP3-containing medium promoted the AR in a high proportion of capacitated spermatozoa (48.6 +/- 3.2%; P < 0.01) compared with control (no rhZP3) samples (14.8 +/- 2.1%). However, rhZP3-containing medium did not cause increased acrosomal exocytosis in non-capacitated spermatozoa (16.8 +/- 3.0%). Induction of the AR was associated with increased tyrosine phosphorylation of a 95 +/- 5 kDa epitope only in capacitated spermatozoa. A dose-dependent increase in the protein phosphorylation of a 95 kDa epitope in response to rhZP3 was detected by [gamma-32P]-ATP labelling of detergent-solubilized sperm proteins. When spermatozoa were co-incubated with monoclonal antibody 97.25 (mAb 97.25) recognizing a 95 kDa tyrosine kinase epitope, there was no rhZP3 induction of tyrosine phosphorylation of the 95 kDa protein. Such co-incubation also markedly inhibited the AR (23.9 +/- 3.1%). These results support the model that initial interaction of the fertilizing spermatozoon with ZP3 involves the tyrosine phosphorylation of a 95 kDa tyrosine kinase protein and that this requires capacitation.

Original languageEnglish
Pages (from-to)1136-44
Number of pages9
JournalMolecular Human Reproduction
Volume4
Issue number12
DOIs
Publication statusPublished - Dec 1998

Keywords

  • Acrosome Reaction/drug effects
  • Animals
  • Antibodies, Monoclonal/pharmacology
  • Blotting, Western
  • Cricetinae
  • Dose-Response Relationship, Drug
  • Egg Proteins/genetics
  • Epitopes
  • Humans
  • Male
  • Membrane Glycoproteins/genetics
  • Phosphorylation
  • Receptors, Cell Surface
  • Recombinant Proteins/genetics
  • Spermatozoa/drug effects
  • Tyrosine/metabolism
  • Zona Pellucida/metabolism
  • Zona Pellucida Glycoproteins

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