Jump to main content
Jump to site search

Issue 23, 2018
Previous Article Next Article

Toward universal protein post-translational modification detection in high throughput format

Author affiliations

Abstract

Post-translational modification (PTM) of proteins plays essential regulatory roles in a variety of pathological conditions. Reliable and practical assays are required to accelerate the discovery of inhibitors and activators for PTM related diseases. Today, methodologies are based on specific or group-specific PTM recognition of e.g. phosphate for kinase activity without extending to other type of PTMs. Here we have established a universal time-resolved luminescence assay on a peptide-break platform for the direct detection of wide variety of PTMs. The developed assay is based on the leucine zipper concept wherein a europium-chelate labeled detection peptide and a non-labeled peptide substrate form a highly luminescent dimer. As an active PTM enzyme at sub or low nanomolar concentration modifies the substrate peptide, the luminescent signal of the detached detection peptide is quenched in the presence of soluble quenchers. The functionality of this universal assay technique has been demonstrated for the monitoring of phosphorylation, dephosphorylation, deacetylation, and citrullination with high applicability also to other PTMs in a high throughput format.

Graphical abstract: Toward universal protein post-translational modification detection in high throughput format

Back to tab navigation

Supplementary files

Publication details

The article was received on 14 Dec 2017, accepted on 30 Jan 2018 and first published on 02 Mar 2018


Article type: Communication
DOI: 10.1039/C7CC09575A
Citation: Chem. Commun., 2018,54, 2910-2913
  • Open access: Creative Commons BY license
  •   Request permissions

    Toward universal protein post-translational modification detection in high throughput format

    H. Härmä, N. Tong-Ochoa, A. J. van Adrichem, I. Jelesarov, K. Wennerberg and K. Kopra, Chem. Commun., 2018, 54, 2910
    DOI: 10.1039/C7CC09575A

    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