Electrophoretic and zymographic characterization of proteins isolated by various extraction methods from ejaculated and capacitated boar sperms
Language English Country Germany Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Electrophoresis, Gel, Two-Dimensional methods MeSH
- Ejaculation MeSH
- Electrophoresis, Polyacrylamide Gel methods MeSH
- Glycoproteins analysis isolation & purification metabolism MeSH
- Hyaluronoglucosaminidase analysis metabolism MeSH
- Sperm Capacitation MeSH
- Peptide Hydrolases analysis metabolism MeSH
- Proteins analysis isolation & purification metabolism MeSH
- Spermatozoa chemistry enzymology MeSH
- Sus scrofa physiology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Glycoproteins MeSH
- Hyaluronoglucosaminidase MeSH
- Peptide Hydrolases MeSH
- Proteins MeSH
The presented work focuses on electrophoretic and zymographic characterization of boar sperm proteins isolated by various extraction methods and on comparison of the protein profiles obtained from ejaculated and in vitro capacitated spermatozoa. Sperm proteins of ejaculated and in vitro capacitated boar sperms were isolated with the following agents: 1% v/v Triton X-100, 1% v/v Triton X-114, 2% v/v acetic acid, 1% m/v sodium dodecyl sulphate (SDS), 30 mM N-octyl-β-D-glucopyranoside (OBG), rehydration buffer (RHB) for isoelectric focusing and finally by the freezing-thawing approach. The extracts were characterized in terms of 1-DE, 2-DE protein profiles, 1-DE glycoprotein staining and proteinase and hyaluronidase substrate zymographic profiles. The results have shown quantitative and qualitative differences in 1-DE protein and glycoprotein profiles with respect to the employed isolation approach. These differences were seen even more clearly in 2-DE protein profiles, where it was possible to distinguish the presence/absence, changes in relative abundance and pI/M(r) shifts of various protein spots. Proteinase and hyaluronidase zymograms supported the prediction that various isolation protocols result in various profiles of enzymatically active molecules.
References provided by Crossref.org
Kinetic Study of 17α-Estradiol Mechanism during Rat Sperm Capacitation
New Insight into Sperm Capacitation: A Novel Mechanism of 17β-Estradiol Signalling
