Investigation on PLK2 and PLK3 substrate recognition
Investigation on PLK2 and PLK3 substrate recognition
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DOI：
10.1016/j.bbapap.2012.07.003
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发表时间：
2012-12-01
影响因子：
3.2
通讯作者：
Pinna, L. A.
中科院分区：
文献类型：
--
作者：
Salvi, M.;Trashi, E.;Pinna, L. A.
基于磷酸Ser/Thr周围氨基酸一级结构的分析表明，相当大比例的亚磷酸盐是由有限数量的嗜酸性激酶产生的，其中蛋白激酶CK2起着突出的作用。最近，属于Polo样激酶家族的新的嗜酸性蛋白激酶已被鉴定，特别涉及PLK1、PLK2和PLK3蛋白。虽然在破译PLK1依赖的磷酸蛋白质组方面已经取得了一些进展，但对PLK2和PLK3激酶的靶标知之甚少。在这篇报道中，我们使用了一种体外方法，包括细胞裂解产物的磷酸化，2D凝胶电泳法和质谱法分离磷酸蛋白，我们描述了PLK2和PLK3激酶的新的潜在底物的鉴定。我们已经鉴定并验证为PLK2和PLK3的体外底物HSP90。GRP-94、β-微管蛋白、钙粘蛋白和14-3-3 epsilon。PLK3在这些蛋白质中产生的亚磷酸盐已经通过质谱分析进行了鉴定，以获得对PLKs特异性决定因素的新见解。后者已被PLKs底物结合区的电子分析进一步证实。(C)2012爱思唯尔B.V.保留所有权利。
Analyses of human phosphoproteome based on primary structure of the aminoacids surrounding the phosphor Ser/Thr suggest that a significant proportion of phosphosites is generated by a restricted number of acidophilic kinases, among which protein kinase CK2 plays a prominent role. Recently, new acidophilic kinases belonging to the Polo like kinase family have been characterized, with special reference to PLK1, PLK2, and PLK3 kinases. While some progress has been made in deciphering the PLK1-dependent phosphoproteome, very little is known about the targets of PLK2 and PLK3 kinases. In this report by using an in vitro approach, consisting of cell lysate phosphorylation, phosphoprotein separation by 2D gel electrophoresis and mass spectrometry, we describe the identification of new potential substrates of PLK2 and PLK3 kinases. We have identified and validated as in vitro PLK2 and PLK3 substrates HSP90. GRP-94, beta-tubulin, calumenin, and 14-3-3 epsilon. The phosphosites generated by PLK3 in these proteins have been identified by mass spectrometry analysis to get new insights about PLKs specificity determinants. These latter have been further corroborated by an in silico analysis of the PLKs substrate binding region. (C) 2012 Elsevier B.V. All rights reserved.