"Purification of maturation-promoting factor, an intracellular regulato" by Manfred J. Lohka, Marianne K. Hayes et al.

Veterinary and Biomedical Sciences, Department of

 

Document Type

Article

Date of this Version

1988

Comments

Proc. Natl. Acad. Sci. USA Vol. 85, pp. 3009-3013, May 1988. Used by permission

Abstract

Maturation-promoting factor causes germinal vesicle breakdown when injected into Xenopus oocytes and can induce metaphase in a cell-free system. The cell-free assay was used to monitor maturation-promoting factor during its purification from unfertilized Xenopus eggs. Ammonium sulfate precipitation and six chromatographic procedures resulted in a preparation purified >3000-fold that could induce germinal vesicle breakdown within 2 hr when injected into cycloheximide- treated oocytes. Proteins of 45 kDa and 32 kDa were correlated with fractions of highest activity in both assays. These fractions contained a protein kinase activity able to phosphorylate the endogenous 45-kDa protein, as well as histone Hi, phosphatase inhibitor 1, and casein. The highly purified preparations described here should help to identify the mechanism of action of maturation-promoting factor and to elucidate the role of protein kinases in the induction of metaphase.

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