Effects of protein kinase inhibitors on the progression of the cell cycle. especially on the initiation of M phase.

蛋白激酶抑制剂对细胞周期进程的影响。

• 批准号: 03670093
• 负责人: YAMASHITA Shigeru
• 金额: $ 1.28万
• 依托单位: Toho University (1992)The University of Tokyo (1991)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670093/
• 关键词: Cell cycle; Initiation of M phase; MPF; Cdc2 kinase; Protein kinase inhibitors; プロテインキナーゼ阻害棄; cdc2キナ-ゼ; プロテインキナ-ゼ

Ammonium sulfate fraction of maturation-promoting factor (MPF) was prepared from unfertilized eggs of Xenopus laevis. The inhibitory effects of K252 derivatives and staurosporine on MPF and cdc2 Kinase activities of the MPF fraction were examined. K252a and staurosporine inhibited the MPF activity with IC50 of 15 and 1.5 ug/ml, respectively and the cdc2 Kinase activity with IC50 of 3.5 and 0.4 ug/ml, respectively. K252b and KT5926 inhibited the cdc2 kinase activity with IC50 of 0.15 and 60 ug/ml, respectively. The decreasing order of the protein kinase inhibitors in the potency of the inhibitory actions was K252b> staurosporine> K252a> KT5926. Thus, the property of cdc2 kinase is remarkably different from that of the other protein kinases, especially in that its sensitivity to K252b is greater by more than 20 times than to K252a. The effects of the protein kinase inhibitors on the processes leading to the activation of MPF were also examined. Although 30 ug/ml K252a and 1.5 ug/ml K252b inhibited cdc2 kinase approximately to the same extent, the former inhibited the dephosphorylation of cdc2 protein and the thiophosphorylation of p110, whereas the latter did not. Therefore, the protein kinase responsible for the activation of MPF is suggeted to be different from cdc2 kinase itself. As to the inhibitory effects on the initiation of M phase of cultured cells staurosporine was the strongest, followed by K252a and K252b. This may be due to the low permeability of K252b across the cell membrane, but it is also possible that the inhibitors act on other targets than cdc2 kinase itself.

硫酸铵的成熟因子（MPF）是由未施用的Xenopus laevis卵制备的。研究了K252衍生物和星形孢菌素对MPF馏分MPF和CDC2激酶活性的抑制作用。 K252a和星形孢菌素分别以15 ug/ml的IC50抑制MPF活性，以及​​IC50分别为3.5和0.4 ug/ml的Cdc2激酶活性。 K252B和KT5926分别以0.15和60 ug/mL抑制CDC2激酶活性。在抑制作用效力中，蛋白激酶抑制剂的降低是K252B> Staurosporine> K252A> KT5926。因此，Cdc2激酶的特性与其他蛋白激酶的特性大不相同，尤其是因为它对K252B的敏感性比对K252a的敏感性大20倍以上。还检查了蛋白激酶抑制剂对导致MPF激活的过程的影响。尽管30 ug/mL K252a和1.5 ug/ml K252b抑制了CDC2激酶，但前者抑制了Cdc2蛋白的去磷酸化和p110的硫代磷酸化，而后者则没有。因此，负责MPF激活的蛋白激酶与CDC2激酶本身不同。至于对培养细胞的M期的抑制作用最强，其次是K252A和K252B。这可能是由于K252B在细胞膜上的渗透性较低，但抑制剂也可能对其他靶标的Cdc2激酶本身作用。

项目成果

Ohmi, K., Yamashita, S., Sakurai, T. and Nonomura, Y.: "Growth and cytoskeletal proteins of cultured bovine carotid smooth muscle cells." Japan. J. Pharmacol.,. 58, (suppl.I). B8 ((1992))

Ohmi, K.、Yamashita, S.、Sakurai, T. 和 Nonomura, Y.：“培养的牛颈动脉平滑肌细胞的生长和细胞骨架蛋白。”

Ohmi,K., Yamashita,S., Y.and Noromura Y.: "Functions of K252a-induced giant endothelial cells" Japan.J.Pharmacol. 58,(suppl.I). 338 (1992)

Ohmi,K.、Yamashita,S.、Y. 和 Noromura Y.：“K252a 诱导的巨内皮细胞的功能”Japan.J.Pharmacol。

Ohmi,K.,Yamashita,S.,Hashimoto,Y.and Nonomura Y.: "Functions of K252a-induced giant endothelial cells in culture." Japan.J.Pharmacol.,58,(suppl.I),. 58. 338 (1992)

Ohmi,K.、Yamashita,S.、Hashimoto,Y. 和 Nonomura Y.：“培养中 K252a 诱导的巨内皮细胞的功能”。

Ohni, K., Yamashita, S., Hashimoto, Y. and Nonomura Y.: "Functions of K252a‐induced giant endothelial cells in culture." Japan. J. Pharmacol.,. 58, (suppl.I). 338 ((1992))

Ohni, K.、Yamashita, S.、Hashimoto, Y. 和 Nonomura Y.：“培养中 K252a 诱导的巨内皮细胞的功能”，J. Pharmacol.，（增刊 338）。 (1992))

Ohmi,K.,Yamashita,S.,Sakurai,T.and Nonomura,Y.: "Growth and cytoskeletal proteins of cultured bovine carotid smooth muscle cells." Japan.J.Pharmacol.,58,(suppl.I),. 58. 138 (1992)

Ohmi,K.、Yamashita,S.、Sakurai,T. 和 Nonomura,Y.：“培养的牛颈动脉平滑肌细胞的生长和细胞骨架蛋白。”