Discovery of a potential allosteric ligand binding site in CDK2.
Discovery of a potential allosteric ligand binding site in CDK2.
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DOI：
10.1021/cb100410m
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发表时间：
2011-05-20
影响因子：
4
通讯作者：
Schoenbrunn, Ernst
中科院分区：
文献类型：
--
作者：
Betzi, Stephane;Alam, Riazul;Martin, Mathew;Lubbers, Donna J.;Han, Huijong;Jakkaraj, Sudhakar R.;Georg, Gunda I.;Schoenbrunn, Ernst
Cyclin-dependent kinases (CDKs) are key regulatory enzymes in cell cycle progression and transcription. Aberrant activity of CDKs has been implicated in a number of medical conditions, and numerous small molecule CDK inhibitors have been reported as potential drug leads. However, these inhibitors exclusively bind to the ATP site, which is largely conserved among protein kinases, and clinical trials have not resulted in viable drug candidates, attributed in part to the lack of target selectivity. CDKs are unique among protein kinases, as their functionality strictly depends on association with their partner proteins, the cyclins. In an effort to identify potential target sites for disruption of the CDK-cyclin interaction, we probed the extrinsic fluorophore 8-anilino-1-naphthalene sulfonate (ANS) with human CDK2 and cyclin A using fluorescence spectroscopy and protein crystallography. ANS interacts with free CDK2 in a saturation-dependent manner with an apparent Kd of 37 μM, and cyclin A displaced ANS from CDK2 with an EC50 value of 0.6 μM. Co-crystal structures with ANS alone and in ternary complex with ATP site-directed inhibitors revealed two ANS molecules bound adjacent to one another, away from the ATP site, in a large pocket that extends from the DFG region above the C-helix. Binding of ANS is accompanied by substantial structural changes in CDK2, resulting in a C-helix conformation that is incompatible for cyclin A association. These findings indicate the potential of the ANS binding pocket as a new target site for allosteric inhibitors disrupting the interaction of CDKs and cyclins.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1016/j.bbapap.2009.10.002
发表时间：
2010-03-01
影响因子：
3.2
作者：
Echalier, Aude;Endicott, Jane A.;Noble, Martin E. M.
通讯作者：
Noble, Martin E. M.
DOI：
10.1107/s0907444998003254
发表时间：
1998-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Brunger, AT;Adams, PD;Warren, GL
通讯作者：
Warren, GL
影响因子：
2.9
作者：
ADAMS, JA;MCGLONE, ML;TAYLOR, SS
通讯作者：
TAYLOR, SS
影响因子：
4.8
作者：
Cheng, Kin-Yip;Noble, Martin E. M.;Johnson, Louise N.
通讯作者：
Johnson, Louise N.