Work

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

Year: 2018

Type: article

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.

Source: Chemical Communications

DOI: 10.1039/c7cc09575a

Authors: Harri Härmä, Natalia Tong-Ochoa, Arjan J. van Adrichem, Ilian Jelesarov, Krister Wennerberg, Kari Kopra

Institutions: University of Turku, Institute for Molecular Medicine Finland, University of Zurich

Language: English

Topic: Advanced Biosensing Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $1,536

Funder: Luonnontieteiden ja Tekniikan Tutkimuksen Toimikunta