High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages.
High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages.
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DOI：
10.1002/pmic.201400431
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发表时间：
2015-09
期刊：
影响因子：
3.4
通讯作者：
Trost M
中科院分区：
文献类型：
--
作者：
Guo M;Härtlova A;Dill BD;Prescott AR;Gierliński M;Trost M
巨噬细胞是重要的免疫细胞，通过吞噬作用吸收外来颗粒和微生物，在先天免疫的最前沿发挥作用。RAW 264.7细胞系通常用于巨噬细胞和吞噬作用领域的实验。然而，很少有人知道它的功能与初级巨噬细胞相比。在这里，我们通过定量超过2500种吞噬体蛋白，对RAW 264.7和骨髓源性巨噬细胞的吞噬体进行了深入的蛋白质组学表征。我们的数据表明，大量蛋白质存在显著差异，包括重要的受体，如甘露糖受体1和Siglec-1。此外，骨髓来源的巨噬细胞吞噬体通过与内体和溶酶体融合而相当快地成熟，我们使用荧光吞噬测定法验证了这一点。我们为该领域的研究人员提供了宝贵的资源，并建议在研究吞噬体功能时谨慎使用RAW 264.7细胞系。所有MS数据均已保存在ProteomeXchange中，标识符为PXD 001293（http：//proteomecentral.proteomexchange.org/soft/PXD 001293）。
Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in‐depth proteomics characterization of phagosomes from RAW 264.7 and bone marrow derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicate that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec‐1. Moreover, bone marrow derived macrophages phagosomes mature considerably faster by fusion with endosomes and the lysosome which we validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. All MS data have been deposited in the ProteomeXchange with identifier PXD001293 (http://proteomecentral.proteomexchange.org/dataset/PXD001293).