Conversion of SB 203580-insensitive MBP kinase family members to drug-sensitive forms by a single amino-acid substitution
Conversion of SB 203580-insensitive MBP kinase family members to drug-sensitive forms by a single amino-acid substitution
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DOI：
10.1016/s1074-5521(98)90170-3
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发表时间：
1998-06-01
影响因子：
--
通讯作者：
Goedert, M
中科院分区：
文献类型：
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作者：
Eyers, PA;Craxton, M;Goedert, M
背景资料：蛋白激酶的特异性抑制剂具有很大的治疗潜力，但其特异性的分子基础仅知之甚少。我们研究了属于一类吡啶基咪唑类药物SE 203580，其抑制应激活化蛋白（SAP）激酶SAPK 2a/p38和SAPK 2b/p38 β 2，但不抑制其他丝裂原活化蛋白激酶家族成员。与其他蛋白激酶抑制剂一样，SE 203580与SAP激酶2a/p38的ATP结合口袋结合。结果：SAP激酶SAPK 1 γ/JNK 1、SAPK 3和SAPK 4不受SE 203580的抑制，因为它们在SAP激酶2a/p30和SAP激酶2b/p38 β 2的ATP结合区中与Thr 106相当的位置具有甲硫氨酸。使用5个SAP激酶和I型和II型TGF β受体的定点诱变，我们已经确定，对于要被SE 203580抑制的蛋白激酶，该残基的侧链必须不大于苏氨酸的侧链。当侧链甚至更小时，如丝氨酸、丙氨酸或甘氨酸中，对SE 203580抑制的敏感性大大增强。因此，I型TGF β受体（其在相当于SAPK 2a/p38和SAPK 2b/p38 β 2的Thr 106的位置处具有丝氨酸）被SE 203580抑制。这些发现解释了靶向ATP结合位点的药物如何特异性抑制蛋白激酶，并且显示在与SAPK 2a/p38和SAPK 2b/p38的Thr 106相当的位置处存在苏氨酸或更小的氨基酸，p38 β 2可诊断蛋白激酶是否对吡啶基咪唑类抑制剂敏感。
Background: Specific inhibitors of protein kinases have great therapeutic potential, but the molecular basis underlying their specificity is only poorly understood, We have investigated the drug SE 203580 which belongs to a class of pyridinyl imidazoles that inhibits the stress-activated protein (SAP) kinases SAPK2a/p38 and SAPK2b/p38 beta 2 but not other mitogen-activated protein kinase family members. Like inhibitors of other protein kinases, SE 203580 binds in the ATP-binding pocket of SAPK2a/p38.Results: The SAP kinases SAPK1 gamma/JNK1, SAPK3 and SAPK4 are not inhibited by SE 203580, because they have methionine in the position equivalent to Thr106 in the ATP-binding region of SAPK2a/p30 and SAPK2b/p38 beta 2. Using site-directed mutagenesis of five SAP kinases and the type I and type II TGF beta receptors, we have established that for a protein kinase to be inhibited by SE 203580, the sidechain of this residue must be no larger than that of threonine, Sensitivity to inhibition by SE 203580 is greatly enhanced when the sidechain is even smaller, as in serine, alanine or glycine. Thus, the type I TGF beta receptor, which has serine at the position equivalent to Thr106 of SAPK2a/p38 and SAPK2b/p38 beta 2, is inhibited by SE 203580.Conclusions: These findings explain how drugs that target the ATP-binding site can inhibit protein kinases specifically, and show that the presence of threonine or a smaller amino acid at the position equivalent to Thr106 of SAPK2a/p38 and SAPK2b/p38 beta 2 is diagnostic of whether a protein kinase is sensitive to the pyridinyl imidazole class of inhibitor.