title: Identification of cyk, a cyclin B2 kinase, as a novel calcium/calmodulin-dependent protein kinase II and its role during Xenopus laevis oocyte maturation.
abstract: Recently, we have partially purified and characterized a specific cell cycle-regulated cyclin B2 kinase (cyk) from prophase oocytes of Xenopus laevis after an ATP-gamma-S activation step (R. Derua, I. Stevens, E. Waelkens, A. Fernandez, N. Lamb, W. Merlevede, and J. Goris, 1997, Exp. Cell Res. 230, 310-324). In the present paper we describe its purification to homogeneity. We could identify the kinase as a special form of calcium/calmodulin-dependent protein kinase II (CaMKII), consisting of five isoforms with molecular masses ranging from 52 to 83 kDa. At least three of them could be considered as novel. Using an in vivo assay with a synthetic peptide (cyktide), an activation of the kinase was shown at about 50% maturation. Further evidence for this observation came from the injection of the calcium chelator BAPTA and the specific cyk/CaMKII inhibitor AIP. A delay of oocyte maturation of at least 1 h was observed. Besides serine 53, a second cyk phosphorylation site in cyclin B2 was identified as threonine 41. Site-directed mutagenesis of these sites indicated that phosphorylation of these sites in Xenopus cyclin B2 was not required for the hallmark functions of cyclin B2.
authors.affiliation: Afdeling Biochemie, Faculteit Geneeskunde, KULeuven, Herestraat 49, Leuven, B-3000, Belgium.
authors.forename: I
authors.initials: I
authors.lastname: Stevens
authors.forename: R
authors.initials: R
authors.lastname: Derua
authors.forename: E
authors.initials: E
authors.lastname: Rondelez
authors.forename: E
authors.initials: E
authors.lastname: Waelkens
authors.forename: W
authors.initials: W
authors.lastname: Merlevede
authors.forename: J
authors.initials: J
authors.lastname: Goris
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D019926:Cyclin B / Q000378:metabolism*; D005260:Female; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D013379:Substrate Specificity / Q000235:genetics; D014982:Xenopus laevis
chemical_list: D019926:Cyclin B; D017871:Calcium-Calmodulin-Dependent Protein Kinases