title: Biological activity of recombinant human ZP3 produced in vitro: potential for a sperm function test.
abstract: The human zona binding test is the most predictive test of sperm function yet the availability of human zona severely restricts its clinical use. The primary aim of this study was to use a commercially available in-vitro transcription and translation system to produce immobilized recombinant human ZP3 (rhuZP3) on agarose beads. The biological activity of this preparation was examined using sperm binding and the acrosome reaction. Significantly higher levels of sperm binding to rhuZP3 beads (n = 12, P < 0.05) compared with controls were observed and there was a significant induction (n = 12, P < 0.01) in the acrosome reaction after overnight incubation at 37 degrees C in 5% CO2 in air. In conclusion, the in-vitro transcription and translation system can produce sufficient quantities of purified immobilized biologically active rhuZP3. These preliminary experiments will enable further refinements to be made so that a solid-phase sperm function test based on rhuZP3 coated beads is likely to be developed in the near future.
authors.affiliation: Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Whitmarsh
authors.forename: M J
authors.initials: MJ
authors.lastname: Woolnough
authors.forename: H D
authors.initials: HD
authors.lastname: Moore
authors.forename: D P
authors.initials: DP
authors.lastname: Hornby
authors.forename: C L
authors.initials: CL
authors.lastname: Barratt
mesh_terms: D000177:Acrosome / Q000502:physiology; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004527:Egg Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D013094:Spermatozoa / Q000502:physiology*; D014158:Transcription, Genetic; D000072216:Zona Pellucida Glycoproteins
chemical_list: D018076:DNA, Complementary; D004527:Egg Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins