Calcium‐ and Cyclic AMP‐independent, Labile Protein Kinase Appearing during Starfish Oocyte Maturation: its Extraction and Partial Characterization*

Abstract

A burst of protein phosphorylation and an appearance of maturation-promoting factor have been reported to occur shortly before germinal vesicle (nucleus) breakdown (GVBD) in 1-methyladenine-induced oocyte maturation of starfish. To detect if a protein kinase is activated before GVBD, protein kinase activity was compared in maturing oocytes which were just undergoing GVBD and immature oocytes of Asterina pectinifera. The oocytes were homogenized in a buffer modified from that used for extracting amphibian maturation-promoting factor. When the supernatant protein of homogenized immature oocytes was used as a substrate, protein kinase activity in the supernatant of the maturing oocytes was 7-fold higher than that of immature oocytes. The protein kinase in the supernatant of the maturing oocytes showed a high substrate specificity for histone Hl among the exogenous substrates examined, and the activity of the maturing oocytes for histone Hl was 6- to 7-fold higher than that of immature oocytes. The protein kinase detected in the maturing oocytes was very labile and was inhibited neither by ethylene glycol bis(.beta.-aminoethyl ether)N,N,N'',N''-tetraacetic acid nor by the heat-stable inhibitor protein of cyclic AMP-dependent protein kinase. These results indicate that a calcium- and cyclic AMP-independent, labile "maturation-specific protein kinase" appeared before GVBD in maturing oocytes, and suggest its participation in the phosphorylation burst in vivo. The possible correlation of this kinase with maturation-promoting factor and chromosome condensation was discussed.