Correlation of epididymal protease inhibitor Eppin and Semenogelin on human ejaculated spermatozoa.
- Author:
Zeng-Jun WANG
1
;
Hong-Fei WU
;
Li-Xin QIAN
;
Di QIAO
;
Jian-Feng SHAO
;
Wei ZHANG
;
Yuan-Geng SUI
;
Zheng-Quan XU
Author Information
- Publication Type:Journal Article
- MeSH: Humans; Male; Protein Binding; Proteinase Inhibitory Proteins, Secretory; Proteins; chemistry; metabolism; Recombinant Proteins; Seminal Vesicle Secretory Proteins; chemistry; metabolism; Spermatozoa; metabolism
- From: National Journal of Andrology 2006;12(5):428-434
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the correlation of epididymal protease inhibitor(Eppin) and Semenogelin(Sg) on human ejaculated spermatozoa.
METHODSThe experimental approaches include: (1) Immunoprecipitation of Eppin with anti-Eppin from semen; (2) Colocalization of Eppin and Sg by immunofluorescence; (3) Immunoprecipitation of rEppin and rSg;(4) Far-Western blotting of rEppin and rSg;(5) Competition of saturated 125I-rSg binding to rEppin with unlabeled Sg, and direct binding of 125I-rSg to rEppin on a blot; (6) Autoradiography of 125I-rSg with rEppin.
RESULTSEppin-Sg complex present on the surface of human ejaculated spermatozoa, Cys-239 is the only cystein for rEppin binding rSg. Reduction and carboxymethylation of Cys-239 blocks binding of 125I-rEppin to rSg.
CONCLUSIONOur study demonstrates that Eppin and Sg bind to each other on human ejaculated spermatozoa. A disulfide linkage occurs between Sg and Eppin, indicating the specificity of binding.
