Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5
Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5
复制标题
DOI：
10.1111/j.1432-1033.1997.t01-2-00527.x
复制
发表时间：
1997-01-15
期刊：
影响因子：
--
通讯作者：
Moulinoux, JP
中科院分区：
文献类型：
--
作者：
Meijer, L;Borgne, A;Moulinoux, JP
细胞周期蛋白依赖性激酶（cdk）在细胞分裂周期（cdc）的胞内调控中起重要作用。这些激酶及其调节剂在人类肿瘤中经常被解除调节。酶筛选最近导致发现细胞周期蛋白依赖性激酶的特异性抑制剂，如丁内酯I，flavopiridol和嘌呤olomoucine。在一系列C2，N-6，N9-取代的腺嘌呤中，2-（1-乙基-2-羟乙基氨基）-6-苄基氨基-9-异丙基嘌呤（roscovitine）对某些细胞周期蛋白依赖性激酶表现出高效和高选择性。用25种高纯度的激酶（包括蛋白激酶A、G和C亚型、肌球蛋白轻链激酶、酪蛋白激酶2、胰岛素受体酪氨酸激酶、c-src、v-abl）研究roscovitine的激酶特异性。大多数激酶不被roscovitine显著抑制，cdc 2/细胞周期蛋白B、cdk 2/细胞周期蛋白A、cdk 2/细胞周期蛋白E和cdk 5/p35仅被显著抑制（IC 50值分别为0.65、0.7、0.7和0.2 μ M）。cdk 4/细胞周期蛋白D1和cdk 6/细胞周期蛋白D2被roscovitine抑制得很差（IC 50> 100 μ M）。细胞外调节激酶erk 1和erk 2受到抑制，IC 50分别为34 μ M和14 μ M。Roscovitine可逆地阻止海星卵母细胞和海胆胚胎在后期前期。Roscovitine抑制非洲爪蟾卵提取物中的体外M期促进因子活性和体外DNA合成。它能阻断非洲爪蟾卵母细胞的甾体酮诱导的卵母细胞成熟和延长因子eEF-1的体内磷酸化。Roscovitine抑制哺乳动物细胞系的增殖，平均IC 50为16 μ M。在存在Roscovitine的情况下，L1210细胞停滞在G1期，并在G2期积累。roscovitine抑制cdc 2/cyclin B对Ser 55上波形蛋白的体内磷酸化。通过其对某些细胞周期蛋白依赖性激酶的独特选择性，roscovitine为细胞周期研究提供了有用的抗有丝分裂试剂，并可能被证明对控制cdc 2，cdk 2或cdk 5激酶活性失调的细胞感兴趣。
Cyclin-dependent kinases (cdk) play an essential role in the intracellular control of the cell division cycle (cdc). These kinases and their regulators are frequently deregulated in human tumours. Enzymatic screening has recently led to the discovery of specific inhibitors of cyclin-dependent kinases, such as butyrolactone I, flavopiridol and the purine olomoucine. Among a series of C2, N-6, N9-substituted adenines tested on purified cdc2/cyclin B, 2-(1-ethyl-2-hydroxyethylamino)-6-benzylamino-9-isopropylpurine (roscovitine) displays high efficiency and high selectivity towards some cyclin-dependent kinases. The kinase specificity of roscovitine was investigated with 25 highly purified kinases (including protein kinase A, G and C isoforms, myosin light-chain kinase, casein kinase 2, insulin receptor tyrosine kinase, c-src, v-abl). Most kinases are not significantly inhibited by roscovitine, cdc2/cyclin B, cdk2/cyclin A, cdk2/cyclin E and cdk5/p35 only are substantially inhibited (IC50 values of 0.65, 0.7, 0.7 and 0.2 mu M, respectively). cdk4/cyclin D1 and cdk6/cyclin D2 are very poorly inhibited by roscovitine (IC50 > 100 mu M). Extracellular regulated kinases erk1 and erk2 are inhibited with an IC50 of 34 mu M and 14 mu M, respectively. Roscovitine reversibly arrests starfish oocytes and sea urchin embryos in late prophase. Roscovitine inhibits in vitro M-phase-promoting factor activity and in vitro DNA synthesis in Xenopus egg extracts. It blocks progesterone-induced oocyte maturation of Xenopus oocytes and in vivo phosphorylation of the elongation factor eEF-1. Roscovitine inhibits the proliferation of mammalian cell lines with an average IC50 of 16 mu M In the presence of roscovitine L1210 cells arrest in G1 and accumulate in G2. In vivo phosphorylation of vimentin on Ser55 by cdc2/cyclin B is inhibited by roscovitine. Through its unique selectivity for some cyclin-dependent kinases, roscovitine provides a useful antimitotic reagent for cell cycle studies and may prove interesting to control cells with deregulated cdc2, cdk2 or cdk5 kinase activities.