Jump to main content
Jump to site search

Issue 2, 2017
Previous Article Next Article

Label-free target identification using in-gel fluorescence difference via thermal stability shift

Author affiliations

Abstract

Target engagement is a prerequisite for the therapeutic effects of bioactive small molecules, and unbiased identification of their target proteins can facilitate drug discovery and chemical biology research. Structural modifications of bioactive natural products for target identification exhibit potential limitations such as synthetic difficulties, limited supplies from natural sources, and loss of original efficacy. Herein, we developed a label-free method for proteome-wide target identification using in-gel fluorescence difference caused by thermal stability shift, namely TS-FITGE. Quantitative intra-gel image analysis of each protein spot revealed target proteins with shifted thermal stability upon drug engagement, and plotting of melting curves by inter-gel analysis confirmed the positive targets. We demonstrated the robustness and applicability of the TS-FITGE method by identifying target proteins, including membrane-anchored proteins, of complex bioactive compounds. Furthermore, we identified and functionally validated nucleophosmin as a novel target protein of hordenine, a natural product upregulator of in vitro translation.

Graphical abstract: Label-free target identification using in-gel fluorescence difference via thermal stability shift

Back to tab navigation

Supplementary files

Publication details

The article was received on 22 Jul 2016, accepted on 20 Sep 2016 and first published on 22 Sep 2016


Article type: Edge Article
DOI: 10.1039/C6SC03238A
Citation: Chem. Sci., 2017,8, 1127-1133
  • Open access: Creative Commons BY license
  •   Request permissions

    Label-free target identification using in-gel fluorescence difference via thermal stability shift

    H. Park, J. Ha, J. Y. Koo, J. Park and S. B. Park, Chem. Sci., 2017, 8, 1127
    DOI: 10.1039/C6SC03238A

    This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements