Research Of Protein Tyrosine Phosphorylation During Mouse Sperm Capacitation

The increasing of protein tyrosine phosphorylation is an important event during sperm capacitation. The research has shown that tyrosme phosphorylation proteins participate in capacitation through several signal pathways, and directly affect sperm physiological changes such as hyperactive motility and acrosome reaction.Objective: In our research we seperate tyrosine phosphorylation proteins which increasing after capacitation and identify them by MS.Methods: Mouse spermatozoa were capacitated in vitro, then protein tyrosine phosphorylation were detected by Western blot after SDS-PAGE. Two dimension gel electrophoresis was employed to resolve proteins. After analysed by ImageMaster, MALDI-TOF MS was used to identify the proteins.Results: (1) The rate of acrosome reaction induced by A23187 was increasing obviously. It showed that sperm capacitated successfully.(2) Tyrosine phosphorylation of 42,56,66,82 and 95 kDa proteins increasing during capacitation. (3) Eight PMF of protein tyrosine phosphorylation were obstained by MS analysis.Conclusion: Two of eight proteins which identified by MS are unkown. Aconitase 2, Glycerol -3-phosphate dehydrogenase and tektin 2 are novel tyrosine phosphorylation proteins during mouse sperm capacitation.