Endoplasmic reticulum (ER) stress is a convergence point in the cellular pathology of many diseases. In the case of type 2 diabetes and the metabolic syndrome, a major role of ER stress has been recognized in the pathogenic cellular alterations of the main districts involved, such as the adipose tissue, the pancreas and the liver. Mass spectrometry (MS) based shotgun proteomics is widely applied to characterize changes in the proteome between different cellular states and will become an invaluable tool in signal transduction and clinical studies. However, in-depth proteomics analysis has so far required elaborate fractionation and thus extensive MS analysis time or, alternatively, restriction to specific proteins of interest. We recently showed that the near complete yeast proteome can be mapped in single-shot fashion by combining the quadrupole Orbitrap (Q Exactive) with high performance and high resolution chromatography with long columns (Nagaraj et al. MCP 2012). Here we apply our single-shot platform to the dose dependent proteome changes induced in HeLa cells by treatment with tunicamycin. By using a simplified model in a human cell line, we give a comprehensive proteomic description of the dose-dependent features of ER stress. This study aims at revealing putative missing links between the unfolded protein response and cell recovery, the intervention of inflammation and innate immunity and apoptosis.
Single-shot, high resolution and in-depth quantification of endoplasmic reticulum stress in a human cell line
MURGIA, MARTA;
2012
Abstract
Endoplasmic reticulum (ER) stress is a convergence point in the cellular pathology of many diseases. In the case of type 2 diabetes and the metabolic syndrome, a major role of ER stress has been recognized in the pathogenic cellular alterations of the main districts involved, such as the adipose tissue, the pancreas and the liver. Mass spectrometry (MS) based shotgun proteomics is widely applied to characterize changes in the proteome between different cellular states and will become an invaluable tool in signal transduction and clinical studies. However, in-depth proteomics analysis has so far required elaborate fractionation and thus extensive MS analysis time or, alternatively, restriction to specific proteins of interest. We recently showed that the near complete yeast proteome can be mapped in single-shot fashion by combining the quadrupole Orbitrap (Q Exactive) with high performance and high resolution chromatography with long columns (Nagaraj et al. MCP 2012). Here we apply our single-shot platform to the dose dependent proteome changes induced in HeLa cells by treatment with tunicamycin. By using a simplified model in a human cell line, we give a comprehensive proteomic description of the dose-dependent features of ER stress. This study aims at revealing putative missing links between the unfolded protein response and cell recovery, the intervention of inflammation and innate immunity and apoptosis.Pubblicazioni consigliate
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