Inhibition of cyclin-dependent kinases by purine analogues - Crystal structure of human cdk2 complexed with roscovitine
Inhibition of cyclin-dependent kinases by purine analogues - Crystal structure of human cdk2 complexed with roscovitine
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DOI：
10.1111/j.1432-1033.1997.0518a.x
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发表时间：
1997-01-15
期刊：
影响因子：
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通讯作者：
Kim, SH
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文献类型：
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作者：
DeAzevedo, WF;Leclerc, S;Kim, SH
细胞周期蛋白依赖性激酶（cdk）控制细胞分裂周期。这些激酶及其调节剂在人类肿瘤中经常被解除调节。通过筛选纯化的cdc 2/cyclin B上的一系列C2，N-6，N9-取代的腺嘌呤，鉴定了cdks的有效抑制剂roscovitine [2-（1-乙基-2-羟乙基氨基）-6-苄基氨基-9-异丙基嘌呤]。Roscovitine显示出高效率和高选择性（Meijer，L.，Borgne，A.，Mulner，O.，Chong，J.P.J.，吹吧，杰杰，Inagaki，N.，Inagaki，M.，Delcros，J. - G. & Moulinoux，J.- P.（1997）Eur. 243，527-536）。它作为ATP与cdc 2结合的竞争性抑制剂。我们在0.24 nm（2.4埃）分辨率下确定了cdk 2和roscovitine之间的复合物的晶体结构，并细化到R（因子）为0.18。抑制剂的嘌呤部分与cdk 2的腺嘌呤结合口袋结合。抑制剂的苄基环基团的位置使得抑制剂能够与ATP复合物结构中未观察到的酶接触。这个苄基环的位置分析解释了roscovitine抑制cdk 2的特异性。该结构还揭示了roscovitine的（R）-立体异构体与cdk 2结合。（R）-异构体抑制cdc 2/细胞周期蛋白B的效力约为（S）-异构体的两倍。结构/活性研究和cdk 2/roscovitine复合物晶体结构分析的结果应该允许设计更有效的cdk抑制剂。
Cyclin-dependent kinases (cdk) control the cell division cycle (cdc). These kinases and their regulators are frequently deregulated in human tumours. A potent inhibitor of cdks, roscovitine [2-(1-ethyl-2-hydroxyethylamino)-6-benzylamino-9-isopropylpurine], was identified by screening a series of C2,N-6,N9-substituted adenines on purified cdc2/cyclin B. Roscovitine displays high efficiency and high selectivity (Meijer, L., Borgne, A., Mulner, O., Chong, J. P. J., Blow, J. J., Inagaki, N., Inagaki, M., Delcros, J.-G. & Moulinoux, J.-P. (1997) Eur. J. Biochem. 243, 527-536). It behaves as a competitive inhibitor for ATP binding to cdc2. We determined the crystal structure of a complex between cdk2 and roscovitine at 0.24-nm (2.4 Angstrom) resolution and refined to an R(factor) of 0.18. The purine portion of the inhibitor binds to the adenine binding pocket of cdk2. The position of the benzyl ring group of the inhibitor enables the inhibitor to make contacts with the enzyme not observed in the ATP-complex structure. Analysis of the position of this benzyl ring explains the specificity of roscovitine in inhibiting cdk2. The structure also reveals that the (R)-stereoisomer of roscovitine is bound to cdk2. The (R)-isomer is about twice as potent in inhibiting cdc2/cyclin B than the (S)-isomer. Results from structure/activity studies and from analysis of the cdk2/roscovitine complex crystal structure should allow the design of even more potent cdk inhibitors.