PHOSPHORYLATION INDEPENDENT ACTIVATION OF HUMAN CYCLIN-DEPENDENT KINASE-2 BY CYCLIN-A INVITRO
PHOSPHORYLATION INDEPENDENT ACTIVATION OF HUMAN CYCLIN-DEPENDENT KINASE-2 BY CYCLIN-A INVITRO
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DOI：
10.1091/mbc.4.1.79
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发表时间：
1993-01-01
影响因子：
3.3
通讯作者：
HARPER, JW
中科院分区：
文献类型：
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作者：
CONNELLCROWLEY, L;SOLOMON, MJ;HARPER, JW
p33 Cdk 2是一种丝氨酸-苏氨酸蛋白激酶，与细胞周期蛋白A、D和E相关，并参与哺乳动物细胞G1/S转换的控制。最近的证据表明，细胞周期蛋白依赖性激酶2（Cdk 2），像它的同系物Cdc 2，需要细胞周期蛋白结合和磷酸化（苏氨酸-160）在体内激活。然而，Cdc 2和Cdk 2之间激活过程的机械细节的保守程度尚不清楚。我们已经开发了Cdk 2和细胞周期蛋白A的细菌表达和纯化系统，允许在没有细胞提取物的情况下进行激活过程的机械研究。重组Cdk 2作为组蛋白H1激酶基本上是无活性的（
p33Cdk2 is a serine-threonine protein kinase that associates with cyclins A, D, and E and has been implicated in the control of the G1/S transition in mammalian cells. Recent evidence indicates that cyclin-dependent kinase 2 (Cdk2), like its homolog Cdc2, requires cyclin binding and phosphorylation (of threonine-160) for activation in vivo. However, the extent to which mechanistic details of the activation process are conserved between Cdc2 and Cdk2 is unknown. We have developed bacterial expression and purification systems for Cdk2 and cyclin A that allow mechanistic studies of the activation process to be performed in the absence of cell extracts. Recombinant Cdk2 is essentially inactive as a histone H 1 kinase (