Purification and characterization of a p13suc1-bound serine/threonine kinase that is expressed in mature rat brain.
We previously reported that the histone-H1 kinase activity bound to p13suc1 increased dramatically during development of the rat brain. In the present work, an in situ kinase assay in an SDS/polyacrylamide gel that contained substrate proteins was employed to characterize the enzyme. Two major proteins of 45 kDa and 100 kDa were found to have p13suc1-bound histone-H1 kinase activity. The former (p45) exhibited strong activity towards histone H1 and had weak autophosphorylation activity, whereas the latter (p100) acted on myelin basic protein or histone H1, and underwent autophosphorylation. p45 was further purified from the nuclear-enriched fraction of rat brain to near homogeneity through sequential column chromatographies. The purified enzyme retained its ability to bind specifically to p13suc1, which suggests that this binding does not require a cofactor. The immunochemical and enzymatic properties of p45 revealed that it differs from Cdk that are known to bind to p13suc1 with high affinity. However, in vitro p45 acted on the peptide motif that is conserved among substrates for cyclin-dependent kinases (Cdk) and mitogen-activated protein kinases, which implies that this protein might belong to the large family of proline-directed kinases. The evidence obtained in this study suggest that p45 is a nuclear p13suc1-bound kinase that has unique functions in the mature brain.
Tamaru
T
T
Division of Protein Metabolism, Osaka University, Japan.
Okada
M
M
D000180:Acrylic Resins; D000367:Age Factors; D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000254:growth & development; D016203:CDC2 Protein Kinase / Q000378:metabolism; D018797:Cell Cycle Proteins; D005656:Fungal Proteins / Q000378:metabolism*; D017403:In Situ Hybridization / Q000379:methods; D016200:Maturation-Promoting Factor / Q000737:chemistry / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D029702:Schizosaccharomyces pombe Proteins; D013379:Substrate Specificity; D014018:Tissue Distribution; D000099166:Polyacrylamides
D000180:Acrylic Resins; D016203:CDC2 Protein Kinase; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D016200:Maturation-Promoting Factor; D017346:Protein Serine-Threonine Kinases; D029702:Schizosaccharomyces pombe Proteins; D000099166:Polyacrylamides; C051655:Suc1 protein, S pombe