Two-dimensional polyacrylamide gel analysis of changes in protein phosphorylation during maturation of Xenopus...

A three- to five-fold increase in non-cAMP-dependent protein phosphorylation has previously been found to occur in progesterone-treated oocytes shortly before germinal vesicle breakdown (GVBD) or immediately following maturation-promoting factor (MPF) injection. Analysis of phosphoprotein from 32Pi-labeled oocytes by both equilibrium and nonequilibrium two-dimensional gel electrophoresis revealed a large number of qualitative changes in phosphoproteins at GVBD, including both phosphorylation and dephosphorylation events. Time-course studies demonstrated that some of the new phosphoproteins appeared as early as 0.36 GVBD50, and all changes were stable at least through GVBD. The pattern of new phosphoproteins at GVBD was similar in oocytes microinjected with a partially purified preparation of MPF. A number of the new phosphoproteins were heat stable, which may facilitate their purification and characterization. These results support the hypothesis that key regulatory events during oocyte maturation are controlled by protein phosphorylation.