From the ‡Institute of Clinical Microbiology, Immunology and Hygiene, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
Institute for Cell and Molecular Biosciences, University of Newcastle, Newcastle, UK.
Mol Cell Proteomics. 2019 Apr;18(4):669-685. doi: 10.1074/mcp.RA118.000929. Epub 2019 Jan 11.
Immune sensing of relies on recognition by macrophages. Mycobacterial cord factor, trehalose-6,6'-dimycolate (TDM), is the most abundant cell wall glycolipid and binds to the C-type lectin receptor (CLR) MINCLE. To explore the kinase signaling linking the TDM-MINCLE interaction to gene expression, we employed quantitative phosphoproteome analysis. TDM caused upregulation of 6.7% and suppressed 3.8% of the 14,000 phospho-sites identified on 3727 proteins. MINCLE-dependent phosphorylation was observed for canonical players of CLR signaling ( PLCγ, PKCδ), and was enriched for PKCδ and GSK3 kinase motifs. MINCLE-dependent activation of the PI3K-AKT-GSK3 pathway contributed to inflammatory gene expression and required the PI3K regulatory subunit p85α. Unexpectedly, a substantial fraction of TDM-induced phosphorylation was MINCLE-independent, a finding paralleled by transcriptome data. Bioinformatics analysis of both data sets concurred in the requirement for MINCLE for innate immune response pathways and processes. In contrast, MINCLE-independent phosphorylation and transcriptome responses were linked to cell cycle regulation. Collectively, our global analyses show substantial reprogramming of macrophages by TDM and reveal a dichotomy of MINCLE-dependent and -independent signaling linked to distinct biological responses.
依赖于巨噬细胞的识别来感知 。分枝杆菌 cord 因子，海藻糖 -6,6'-二(mycoloyl)（TDM）是最丰富的细胞壁糖脂，与 C 型凝集素受体（CLR）MINCLE 结合。为了探索将 TDM-MINCLE 相互作用与基因表达联系起来的激酶信号通路，我们采用了定量磷酸化蛋白质组学分析。TDM 导致 14000 个磷酸化位点中的 6.7%上调和 3.8%下调。在 CLR 信号的典型参与者（PLCγ、PKCδ）中观察到 MINCLE 依赖性磷酸化，并且富含 PKCδ 和 GSK3 激酶基序。MINCLE 依赖性激活 PI3K-AKT-GSK3 通路有助于炎症基因表达，需要 PI3K 调节亚基 p85α。出乎意料的是，相当一部分 TDM 诱导的磷酸化是 MINCLE 非依赖性的，这一发现与转录组数据相平行。对这两个数据集的生物信息学分析都一致认为，MINCLE 是先天免疫反应途径和过程所必需的。相比之下，MINCLE 非依赖性磷酸化和转录组反应与细胞周期调控有关。总的来说，我们的全面分析表明 TDM 对巨噬细胞进行了大量的重新编程，并揭示了与不同生物学反应相关的 MINCLE 依赖性和非依赖性信号的二分法。
