論文

査読有り 国際誌
2018年12月12日

Chemical Profiling of the Endoplasmic Reticulum Proteome Using Designer Labeling Reagents.

Journal of the American Chemical Society
  • Alma Fujisawa
  • ,
  • Tomonori Tamura
  • ,
  • Yuki Yasueda
  • ,
  • Keiko Kuwata
  • ,
  • Itaru Hamachi

140
49
開始ページ
17060
終了ページ
17070
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/jacs.8b08606
出版者・発行元
AMER CHEMICAL SOC

The endoplasmic reticulum (ER) is an organelle that performs a variety of essential cellular functions via interactions with other organelles. Despite its important role, chemical tools for profiling the composition and dynamics of ER proteins remain very limited because of the labile nature of these proteins. Here, we developed ER-localizable reactive molecules (called ERMs) as tools for ER-focused chemical proteomics. ERMs can spontaneously localize in the ER of living cells and selectively label ER-associated proteins with a combined affinity and imaging tag, enabling tag-mediated ER protein enrichment and identification with liquid chromatography tandem mass spectrometry (LC-MS/MS). Using this method, we performed proteomic analysis of the ER of HeLa cells and newly assigned three proteins, namely, PAICS, TXNL1, and PPIA, as ER-associated proteins. The ERM probes could be used simultaneously with the nucleus- and mitochondria-localizable reactive molecules previously developed by our group, which enabled orthogonal organellar chemoproteomics in a single biological sample. Moreover, quantitative analysis of the dynamic changes in ER-associated proteins in response to tunicamycin-induced ER stress was performed by combining ER-specific labeling with SILAC (stable isotope labeling by amino acids in cell culture)-based quantitative MS technology. Our results demonstrated that ERM-based chemical proteomics provides a powerful tool for labeling and profiling ER-related proteins in living cells.

リンク情報
DOI
https://doi.org/10.1021/jacs.8b08606
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30433779
ID情報
  • DOI : 10.1021/jacs.8b08606
  • ISSN : 0002-7863
  • PubMed ID : 30433779

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