Quantitative phosphoproteomic analysis of IL-33-mediated signaling.
Quantitative phosphoproteomic analysis of IL-33-mediated signaling.
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DOI：
10.1002/pmic.201400303
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发表时间：
2015-01
期刊：
影响因子：
3.4
通讯作者：
Pandey A
中科院分区：
文献类型：
--
作者：
Pinto SM;Nirujogi RS;Rojas PL;Patil AH;Manda SS;Subbannayya Y;Roa JC;Chatterjee A;Prasad TS;Pandey A
白介素33(IL-33)是细胞因子IL-1家族中的一个新成员，在免疫反应的调节中发挥着不同的作用。IL-33通过异二聚体受体复合体发挥作用，导致促炎细胞因子的产生和释放。对IL-33激活的信号通路的详细了解仍不清楚。为了深入了解IL-33介导的信号机制，我们进行了基于SILAC的全球定量磷酸蛋白质组分析，鉴定了来自2,746个蛋白质的7,191个磷酸化位点。我们观察到在IL-33刺激下，对应于672个蛋白质的1,050个位点的磷酸化水平发生了变化。我们首次报道了多种蛋白激酶的磷酸化，包括由IL-33诱导的丝裂原活化蛋白激酶2(Mapkapk2)、受体(TNFRSF)相互作用的丝氨酸苏氨酸激酶1(Ripk1)和NAD激酶(NADK)。此外，我们观察到IL-33诱导的几种蛋白磷酸酶的磷酸化，包括蛋白酪氨酸磷酸酶，非受体类型12(Ptpn12)和肌醇多聚磷酸酶D(INPP5D)，这些都是以前没有报道过的。网络分析显示，肌动蛋白结合和细胞骨架重组的丰富可能在IL-33诱导的巨噬细胞激活中起重要作用。本研究首次对IL-33调节的磷酸蛋白质组进行了定量分析。我们的发现极大地扩展了对IL-33介导的信号事件的理解，并有可能为免疫相关疾病(如哮喘)提供新的治疗靶点，其中观察到IL-33的异常调节。
Interleukin-33 (IL-33) is a novel member of the IL-1 family of cytokines that plays diverse roles in the regulation of immune responses. IL-33 exerts its effects through a heterodimeric receptor complex resulting in the production and release of pro-inflammatory cytokines. A detailed understanding of the signaling pathways activated by IL-33 is still unclear. To gain insights into the IL-33 mediated signaling mechanisms, we carried out a SILAC-based global quantitative phosphoproteomic analysis that resulted in the identification of 7,191 phosphorylation sites derived from 2,746 proteins. We observed alterations in the level of phosphorylation in 1,050 sites corresponding to 672 proteins upon IL-33 stimulation. We report, for the first time, phosphorylation of multiple protein kinases, including Mitogen-activated protein kinase-activated protein kinase 2 (Mapkapk2), Receptor (TNFRSF)-interacting serine-threonine kinase 1 (Ripk1) and NAD kinase (Nadk) that are induced by IL-33. In addition, we observed IL-33-induced phosphorylation of several protein phosphatases including Protein tyrosine phosphatase, Non-receptor type 12 (Ptpn12) and Inositol polyphosphate-5-phosphatase D (Inpp5d), which have not been reported previously. Network analysis revealed an enrichment of actin binding and cytoskeleton reorganization that could be important in macrophage activation induced by IL-33. Our study is the first quantitative analysis of IL-33-regulated phosphoproteome. Our findings significantly expand the understanding of IL-33 mediated signaling events and have the potential to provide novel therapeutic targets pertaining to immune related diseases such as asthma where dysregulation of IL-33 is observed.