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Rapid and sensitive profiling and quantification of the human cell line proteome by LC-MS/MS without prefractionation

Authors

  • Xuefei Yin,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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    • These authors contributed equally to this work.

  • Xiaohui Liu,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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    • These authors contributed equally to this work.

  • Yang Zhang,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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  • Guoquan Yan,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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  • Fang Wang,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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  • Haojie Lu,

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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  • Huali Shen,

    Corresponding author
    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
    • Correspondence: Professor Pengyuan Yang, Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai 200032, P. R. China

      E-mail: pyyang@fudan.edu.cn

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  • Pengyuan Yang

    1. Department of Chemistry and Institutes of Biomedical Sciences Shanghai Medical School, Fudan University, Shanghai, P. R. China
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Abstract

In this paper, we demonstrate a rapid and reproducible 1D LC-MS/MS workflow for fast quantitative proteomic research. We have optimized the LC-MS/MS conditions, including digestion and gradient conditions, sample loading amount, and MS parameter settings. As a result, we were able to obtain twice as many protein identifications compared with the LC-MS/MS conditions before optimization. More than 4500 protein groups and 50 000 peptides were identified in less than 8 h without any fractionation. This 1D workflow was then applied to the analysis of the MLN4924 treated/untreated human umbilical vein endothelial cell (HUVEC) samples with label-free quantification. In these experiments, a total of 179 proteins showed a statistically significant expression change after the MLN4924 treatment. Functional analysis showed that these proteins are associated with cell death and survival; gene expression; cell cycle; and DNA replication, recombination, and repair.

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