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Outlining the Grb2 interactome data and its interacting partners in HEK293 cells in absence and presence of epidermal growth factor

机译:概述在不存在和存在表皮生长因子的情况下HEK293细胞中的Grb2相互作用组数据及其相互作用伙伴

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摘要

Growth factor receptor-bound protein 2 (Grb2) is an adaptor protein involved in the signal transduction pathways. This dataset enlists proteins which interact with Grb2 in the presence and absence of a mitogenic stimulus. Grb2 expressing HEK293 cells were cultured in light and heavy labeled SILAC media. Normal lysine and arginine were incorporated as light labels while 8 and 10 Da heavier labels of respective isotopes were used for heavy labeling. While light labeled cells were used to enrich basal Grb2 interactome, the heavy labeled cells were stimulated in presence of epidermal growth factor (EGF) to investigate the altered Grb2 interactome dynamics. Equal number of EGF stimulated and non-stimulated cells was pooled, lysed and subjected to affinity purification coupled to mass spectrometry (AP-MS). The variety of Grb2 protein partners changed as a consequence of EGF stimulation. Additionally, SILAC labeling helped in quantitative estimation of altered association of a few interactors with the bait protein. Data are available via PRIDE repository with the dataset identifier PXD012957 (https://www.ebi.ac.uk/pride/archive/projects/PXD012957).
机译:生长因子受体结合蛋白2(Grb2)是一种参与信号转导途径的衔接蛋白。该数据集列出了在有丝分裂刺激存在和不存在的情况下与Grb2相互作用的蛋白质。表达Grb2的HEK293细胞在轻度和重度标记SILAC培养基中培养。正常的赖氨酸和精氨酸被掺入作为轻标记,而相应同位素的8和10 Da较重的标记被用于重标记。虽然使用轻标记的细胞富集基础的Grb2相互作用组,但在存在表皮生长因子(EGF)的情况下刺激重标记的细胞,以研究改变的Grb2相互作用组的动力学。合并,裂解等量的EGF刺激和未刺激的细胞,并进行与质谱(AP-MS)偶联的亲和纯化。由于EGF刺激,Grb2蛋白伴侣的种类发生了变化。此外,SILAC标记有助于定量评估一些相互作用物与诱饵蛋白之间的关联改变。可通过PRIDE存储库获得数据集标识符为PXD012957(https://www.ebi.ac.uk/pride/archive/projects/PXD012957)的数据。

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