Identification and characterization of oviductal glycoprotein-binding protein partner on gametes: epitopic similarity to non-muscle myosin IIA, MYH 9
Open Access
- 27 March 2006
- journal article
- Published by Oxford University Press (OUP) in Molecular Human Reproduction
- Vol. 12 (4), 275-282
- https://doi.org/10.1093/molehr/gal028
Abstract
The mammalian estrogen induced oviductal glycoprotein (OGP) has been known to associate with capacitated sperm, oocytes and developing embryos. This study aimed to identify the putative binding partner of OGP on gametes using N-terminal peptide of bonnet monkey (Macaca radiata) OGP, Nmon, as bait. A protein(s) of molecular size ∼54 kDa was detected by far-western blot analysis of detergent solubilized human sperm proteins. MALDI-TOF mass spectra analysis of ∼54 kDa tryptic peptides gave a significant hit to non-muscle myosin heavy chain. Biochemical characterization of ∼54 kDa was done with antibodies specific to non-muscle myosin IIA, MYH9. The ∼54 kDa protein, possible breakdown product of MYH9, immunoreacted with MYH9 antibody in western blot analysis. OGP binding to ∼54 kDa could also be demonstrated in far-western blot analysis of detergent solubilized human sperm proteins and nuclear matrix intermediate filament (NM-IF) preparations from human sperm and mouse oocytes. Far-western blot analysis of MYH9 enriched by immunoprecipitation identified the native ∼220 kDa protein as OGP-binding partner. The identical and characteristic immunogold localization pattern of Nmon and MYH9 on sperm NM-IF preparation substantiated these findings. The results suggest that OGP binds to both gametes through its interaction with MYH9 through the non-glycosylated N-terminal conserved region of OGP, spanning the residues 11–137.Keywords
This publication has 34 references indexed in Scilit:
- Involvement of an F-actin skeleton on the acrosome reaction in guinea pig spermatozoaCell Motility, 2000
- Microfilament stabilization by jasplakinolide arrests oocyte maturation, cortical granule exocytosis, sperm incorporation cone resorption, and cell-cycle progression, but not DNA replication, during fertilization in miceMolecular Reproduction and Development, 2000
- Characteristics of an Oviductal Glycoprotein and its Potential Role in the Fertilization Process1Biology of Reproduction, 1998
- Cortical granule exocytosis in hamster eggs requires microfilamentsMolecular Reproduction and Development, 1997
- Microfilament assembly and cortical granule distribution during maturation, parthenogenetic activation and fertilisation in the porcine oocyteZygote, 1996
- Association of oviduct-specific glycoproteins with human and baboon (Papio anubis) ovarian oocytes and enhancement of human sperm binding to human hemizonae following in vitro incubationBiology of Reproduction, 1996
- Myosin II filament assemblies in the active lamella of fibroblasts: their morphogenesis and role in the formation of actin filament bundles.The Journal of cell biology, 1995
- Identification of a Sperm Penetration Factor in the Oviduct of the Golden Hamster1Biology of Reproduction, 1995
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences of the United States of America, 1979
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970