Issue 23, 2018

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

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

Supplementary files

Article information

Article type
Communication
Submitted
14 Dec 2017
Accepted
30 Jan 2018
First published
02 Mar 2018
This article is Open Access
Creative Commons BY license

Chem. Commun., 2018,54, 2910-2913

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

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