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Effect of Recombinant hTSBP on Activity of PKA and PKC in Human Sperm
Wen-Hui Su, Guo-Li Wang, Jie Zhang, Bo Jin, Chen Feng, Zhe Zhang, Xiao-Yan Xu, Bing-Zhi Yu*
Department of Biochemistry and Molecular Biology, China Medical University, Shenyang 110001, China
Abstract: To investigate the molecular mechanism of promotion of human sperm capacitation by human testis sperm binding protein (hTSBP)， the effects of hTSBP on activity of protein kinase A (PKA) and protein kinase C (PKC) in human sperm were detected. Recombinant plasmid pcDNA3.1/myc-His(-)B-tsbp was used to transfect HEK293 cell and recombinant protein His6-hTSBP was expressed and purified by immobilized metal ion affinity chromatography (IMAC); Human sperm from healthy volunteer was treated with the recombinant protein， then the plasma protein and the membrane protein were extracted respectively. His6-hTSBP was found to combine to human sperm membrane in vitro. The result of autoradiography showed that 0.1 mg/ml His6-hTSBP could increase the activity of PKA in human sperm plasma， but had no significant effect on the activity of PKC in sperm membrane.
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