From the ‡State Key Laboratory of Cellular Stress Biology, Innovation Center for Cellular Signaling Network, School of Life Sciences, Xiamen University.
From the ‡State Key Laboratory of Cellular Stress Biology, Innovation Center for Cellular Signaling Network, School of Life Sciences, Xiamen University
Mol Cell Proteomics. 2019 Jun;18(6):1054-1069. doi: 10.1074/mcp.RA119.001380. Epub 2019 Mar 8.
Lipopolysaccharide (LPS)-induced macrophage activation is a prototype of innate immune response. Although key effector proteins in LPS signaling pathway have been revealed, the map of dynamic protein interactions and phosphorylation as well as the stoichiometry of protein complexes are lacking. Here we present a dynamic map of protein interactions and phosphorylation in MyD88, TRAF6 and NEMO complexes obtained by SWATH-MS. The comprehensive MS measurement leads to quantification of up to about 3,000 proteins across about 21-40 IP samples. We detected and quantified almost all known interactors of MyD88, TRAF6 and NEMO. By analyzing these quantitative data, we uncovered differential recruitment of IRAK family proteins to LPS-induced signaling complexes and determined the stoichiometry of the Myddosome complex. In addition, quantitative phosphoproteomics analysis identified a number of unreported high-confidence phosphosites on the key proteins in LPS signaling pathway. Collectively, data of dynamic protein interactions and phosphorylation presented by this study could be a resource for further study of the LPS signaling pathway.
脂多糖（LPS）诱导的巨噬细胞激活是先天免疫反应的原型。尽管已经揭示了 LPS 信号通路中的关键效应蛋白，但动态蛋白质相互作用和磷酸化图谱以及蛋白质复合物的化学计量学仍然缺乏。在这里，我们通过 SWATH-MS 呈现了 MyD88、TRAF6 和 NEMO 复合物中蛋白质相互作用和磷酸化的动态图谱。全面的 MS 测量可定量约 21-40 个 IP 样本中的多达约 3000 种蛋白质。我们检测和定量了 MyD88、TRAF6 和 NEMO 的几乎所有已知相互作用蛋白。通过分析这些定量数据，我们揭示了 IRAK 家族蛋白在 LPS 诱导的信号复合物中的差异募集，并确定了 Myddosome 复合物的化学计量。此外，定量磷酸蛋白质组学分析鉴定了 LPS 信号通路中关键蛋白的一些未报道的高可信度磷酸化位点。总之，本研究中呈现的动态蛋白质相互作用和磷酸化数据可以成为 LPS 信号通路进一步研究的资源。
