ATP-competitive LRRK2 inhibitors interfere with monoclonal antibody binding to the kinase domain of LRRK2 under native conditions. A method to directly monitor the active conformation of LRRK2?
ATP-competitive LRRK2 inhibitors interfere with monoclonal antibody binding to the kinase domain of LRRK2 under native conditions. A method to directly monitor the active conformation of LRRK2?
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DOI：
10.1016/j.jneumeth.2012.12.015
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发表时间：
2013-03-30
影响因子：
3
通讯作者：
Gloeckner, Christian J.
中科院分区：
文献类型：
--
作者：
Gillardon, Frank;Kremmer, Elisabeth;Gloeckner, Christian J.
富亮氨酸重复激酶2 （LRRK2）突变是帕金森病最常见的遗传原因。LRRK2激酶活性是神经细胞培养毒性所必需的，这表明选择性激酶抑制剂可能预防患者的神经退行性变。直接监测细胞中LRRK2的活性将有利于开发小分子LRRK2抑制剂。在这里，我们证明了针对激活片段的单克隆抗LRRK2抗体在存在atp竞争性LRRK2抑制剂的情况下与天然LRRK2蛋白的结合效率较低。由于激酶抑制剂阻止自磷酸化和激活片段的再折叠，我们假设抗体在天然条件下优先结合LRRK2的活性构象。(c) 2013 Elsevier B.V.版权所有
Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of Parkinson's disease. LRRK2 kinase activity is required for toxicity in neuronal cell cultures suggesting that selective kinase inhibitors may prevent neurodegeneration in patients. Directly monitoring LRRK2 activity in cells would be advantageous for the development of small molecule LRRK2 inhibitors. Here, we demonstrate that a monoclonal anti-LRRK2 antibody directed against the activation segment binds less efficiently to native LRRK2 protein in the presence of ATP-competitive LRRK2 inhibitors. Since kinase inhibitors prevent autophosphorylation and refolding of the activation segment, we hypothesize that the antibody preferentially binds to the active conformation of LRRK2 under native conditions. (c) 2013 Elsevier B.V. All rights reserved.