Identification of Substrates of Protein Kinase C at 1-Cell Fertilized Eggs in Mouse

Abstract: To study regulatory effects of protein kinase C (PKC) in early development of mouse， identification of the PKC substrates at fertilized eggs was carried out with super-ovulation， in vitro fertilization， in vitro phosphorylation and autoradiography. The endogenous protein kinase in fertilized egg lysates was deactivated through multiply freezing and thawing cycles. The sample without purified PKC did not show any autoradiography band， so it served as blank control in this study. The sample of 55 fertilized eggs with partially purified PKC showed 12 bands in autoradiography. While less autoradiography bands were observed in samples containing both purified PKC and more potent PKC inhibitor (H-7 or staurosporin). According to Rf value of proteins， the relative molecular weights of these phosphorylated proteins were determined as 120 kDa， 100 kDa， 79 kDa， 63 kDa， 59 kDa， 47 kDa， 40 kDa， 34 kDa， 32 kDa， 26 kDa， 24 kDa and 22 kDa， respectively. These results suggested that PKC played an important role in early development of mouse.

CSTR: 32200.14.cjcb.2005.05.0023