Specific CLK inhibitors from a novel chemotype for regulation of alternative splicing.
Specific CLK inhibitors from a novel chemotype for regulation of alternative splicing.
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DOI：
10.1016/j.chembiol.2010.11.009
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发表时间：
2011-01-28
影响因子：
--
通讯作者：
Knapp S
中科院分区：
文献类型：
--
作者：
Fedorov O;Huber K;Eisenreich A;Filippakopoulos P;King O;Bullock AN;Szklarczyk D;Jensen LJ;Fabbro D;Trappe J;Rauch U;Bracher F;Knapp S
越来越多的人认识到蛋白激酶在控制可变剪接中的重要性。为了确定潜在的调节机制，需要高度选择性的抑制剂。在这里，我们报告的二氯吲哚烯氨基腈KH-CB 19，一个有效的和高度特异性的抑制剂的CDC 2样激酶亚型1和4（CLK 1/CLK 4）的发现和表征。KH-CB 19与CLK 1和CLK 3的共晶体结构显示非ATP模拟结合模式，螺旋αC和磷酸结合环的构象变化以及与激酶铰链区的卤素键合。KH-CB 19有效抑制细胞中SR（丝氨酸/精氨酸）蛋白的磷酸化，与其预期的作用机制一致。CLK 1/CLK 4的化学抑制产生剪接因子去磷酸化的独特模式，并且在低nM浓度下对两种组织因子同种型flTF（全长TF）和asHTF（选择性剪接的人TF）的剪接具有深远影响。KH-CB 19共晶体结构揭示了ATP竞争性而非ATP模拟结合模式，KH-CB 19与激酶铰链区形成卤键，KH-CB 19导致SR蛋白的去磷酸化，并影响TF异构体在细胞中的剪接
There is a growing recognition of the importance of protein kinases in the control of alternative splicing. To define the underlying regulatory mechanisms, highly selective inhibitors are needed. Here, we report the discovery and characterization of the dichloroindolyl enaminonitrile KH-CB19, a potent and highly specific inhibitor of the CDC2-like kinase isoforms 1 and 4 (CLK1/CLK4). Cocrystal structures of KH-CB19 with CLK1 and CLK3 revealed a non-ATP mimetic binding mode, conformational changes in helix αC and the phosphate binding loop and halogen bonding to the kinase hinge region. KH-CB19 effectively suppressed phosphorylation of SR (serine/arginine) proteins in cells, consistent with its expected mechanism of action. Chemical inhibition of CLK1/CLK4 generated a unique pattern of splicing factor dephosphorylation and had at low nM concentration a profound effect on splicing of the two tissue factor isoforms flTF (full-length TF) and asHTF (alternatively spliced human TF). ► We report a highly selective nanomolar inhibitor KH-CB19 for the kinases CLK1/4 and DYRK1 ► KH-CB19 cocrystal structures revealed an ATP competitive but not ATP mimetic binding mode ► KH-CB19 formed halogen bonds with the kinase hinge region ► KH-CB19 led to dephosphorylation of SR proteins and effected splicing of TF isoforms in cells
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DOI：
10.1016/j.str.2008.12.023
发表时间：
2009-03-11
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Bullock AN;Das S;Debreczeni JE;Rellos P;Fedorov O;Niesen FH;Guo K;Papagrigoriou E;Amos AL;Cho S;Turk BE;Ghosh G;Knapp S
通讯作者：
Knapp S
影响因子：
4.8
作者：
Jiang, Kun;Patel, Niketa A.;Cooper, Denise R.
通讯作者：
Cooper, Denise R.
影响因子：
5.6
作者：
Doudou, Slimane;Sharma, Raman;Burton, Neil A.
通讯作者：
Burton, Neil A.
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：
10.1073/pnas.0708800104
发表时间：
2007-12-18
影响因子：
11.1
作者：
Fedorov, Oleg;Marsden, Brian;Knapp, Stefan
通讯作者：
Knapp, Stefan