Castillo-Montoya Javier, Ghosh Indraneel
Department of Chemistry and Biochemistry, University of Arizona, 1306 East University Boulevard, Tucson, AZ, 85721, USA.
Methods Mol Biol. 2017;1596:307-319. doi: 10.1007/978-1-4939-6940-1_19.
The over 500 human protein kinases are estimated to phosphorylate at least one-third of the proteome. This posttranslational modification is of paramount importance to intracellular signaling and its deregulation is linked to numerous diseases. Deciphering the specific cellular role of a protein kinase of interest remains challenging given their structural similarity and potentially overlapping activity. In order to exert control over the activity of user-defined kinases and allow for understanding and engineering of complex signal transduction pathways, we have designed ligand inducible split protein kinases. In this approach, protein kinases are dissected into two fragments that cannot spontaneously assemble and are thus inactive. The two kinase fragments are attached to chemical inducers of dimerization (CIDs) that allow for ligand induced heterodimerization and concomitant activation of kinase activity.
据估计，超过500种人类蛋白激酶可使至少三分之一的蛋白质组发生磷酸化。这种翻译后修饰对细胞内信号传导至关重要，其失调与多种疾病相关。鉴于蛋白激酶结构相似且活性可能重叠，解读特定目标蛋白激酶的具体细胞作用仍具有挑战性。为了控制用户定义激酶的活性，并有助于理解和构建复杂的信号转导通路，我们设计了配体诱导型分裂蛋白激酶。在这种方法中，蛋白激酶被分解成两个不能自发组装的片段，因此没有活性。这两个激酶片段与二聚化化学诱导剂（CID）相连，该诱导剂可实现配体诱导的异源二聚化并伴随激酶活性的激活。
