RNA aptamers as pathway-specific MAP kinase inhibitors.
RNA aptamers as pathway-specific MAP kinase inhibitors.
复制标题
DOI：
10.1016/s1074-5521(00)00032-6
复制
发表时间：
2000-11
影响因子：
--
通讯作者：
S. Seiwert;T. Nahreini;S. Aigner;N. Ahn;O. Uhlenbeck
中科院分区：
文献类型：
--
作者：
S. Seiwert;T. Nahreini;S. Aigner;N. Ahn;O. Uhlenbeck
背景：在真核细胞中，许多细胞内信号通路都有密切相关的丝裂原活化蛋白激酶（MAPK）作为中心成分。尽管MAPKs是控制这些信号通路激活所引起的细胞反应的明显靶标，但开发针对涉及MAPKs的特定细胞信号通路的抑制剂已被证明是困难的。结果：我们使用RNA组合方法分离出抑制细胞外调节激酶2 （ERK2）体外磷酸化活性的RNA。这些抑制剂阻断ERK1和ERK2的磷酸化，但不抑制Jun n -末端激酶或p38 MAPKs。动力学分析表明，这些抑制剂通过位阻底物和ATP结合在高皮摩尔浓度下起作用。在一个案例中，我们发现了一个紧凑的RNA结构域负责抑制。结论：RNA试剂可选择性识别和抑制参与单一信号转导通路的MAPKs。这里描述的方法很容易推广，并可用于开发参与其他信号转导途径的mapk抑制剂。这些试剂可能是分析和区分调节不同信号反应的同源效应物的有价值的工具。
Background:In eukaryotic cells, many intracellular signaling pathways have closely related mitogen activated protein kinase (MAPK) paralogs as central components. Although MAPKs are therefore obvious targets to control the cellular responses resulting from the activation of these signaling pathways, the development of inhibitors which target specific cell signaling pathways involving MAPKs has proven difficult.Results:We used an RNA combinatorial approach to isolate RNAs that inhibit the in vitro phosphorylation activity of extracellular regulated kinase 2 (ERK2). These inhibitors block phosphorylation by ERK1 and ERK2, but do not inhibit Jun N-terminal kinase or p38 MAPKs. Kinetic analysis indicates these inhibitors function at high picomolar concentrations through the steric exclusion of substrate and ATP binding. In one case, we identified a compact RNA structural domain responsible for inhibition.Conclusions:RNA reagents can selectively recognize and inhibit MAPKs involved in a single signal transduction pathway. The methodology described here is readily generalizable, and can be used to develop inhibitors of MAPKs involved in other signal transduction pathways. Such reagents may be valuable tools to analyze and distinguish homologous effectors which regulate distinct signaling responses.