Changes in proteasome activities were observed during starfish oocyte maturation induced by a maturation-inducing hormone, 1-methyladenine. Succinyl-Leu-Leu-Val-Tyr-MCA-hydrolyzing proteasome activity in immature oocytes showed a main peak of a 1500-kDa fraction and a shoulder centered at a 650-kDa fraction on Superose 6 gel-filtration chromatography in the presence of ATP and glycerole. The 1500-kDa activity transiently decreased and then increased at about a half the time required for germinal vesicle breakdown (GVBD). In contrast, the 650-kDa activity showed only a slight change during the maturation process. The activity of the 1500-kDa complex, unlike that of the 650-kDa complex, was immunoprecipitated with an antibody raised against regulatory subunits of mammalian 26S proteasomes, whereas both 1500- and 650-kDa activities were immunoprecipitated with anti-20S proteasome antibody. In addition, the 1500-kDa complex showed an ATP/ubiquitin-dependent proteolytic activity. These results indicate that the 1500- and 650-kDa complexes correspond to the mammalian 26S and 20S proteasomes, respectively. Immunoblot analysis revealed that the change in the 26S proteasomal activity is due to the change in the amount of the 20S proteasome subcomplex. Taken together, the proteasome undergoes changes in molecular assembly and activities during hormone-induced oocyte maturation.
Copyright 1999 Academic Press.