Publication details

Analysis of binding interfaces of the human scaffold protein AXIN1 by peptide microarrays

Investor logo
Investor logo
Authors

HARNOŠ Jakub RYNEŠ Jan VÍŠKOVÁ Pavlína TRANTÍRKOVÁ Silvie BAJARD ÉP.ESNER Lola Murielle TRANTÍREK Lukáš BRYJA Vítězslav

Year of publication 2018
Type Article in Periodical
Magazine / Source Journal of Biological Chemistry
MU Faculty or unit

Faculty of Science

Citation
Web http://www.jbc.org/content/293/42/16337
Doi http://dx.doi.org/10.1074/jbc.RA118.005127
Keywords peptide array; scaffold protein; axin; serine; threonine protein kinase; p53; Myc (c-Myc); casein kinase 1E; dishevelled; intrinsically disordered region; Wnt pathway
Description Intrinsically disordered regions (IDRs) are protein regions that lack persistent secondary or tertiary structure under native conditions. IDRs represent >40% of the eukaryotic proteome and play a crucial role in protein-protein interactions. The classical approach for identification of these interaction interfaces is based on mutagenesis combined with biochemical techniques such as coimmunoprecipitation or yeast two-hybrid screening. This approach either provides information of low resolution (large deletions) or very laboriously tries to precisely define the binding epitope via single amino acid substitutions. Here, we report the use of a peptide microarray based on the human scaffold protein AXIN1 for high-throughput and -resolution mapping of binding sites for several AXIN1 interaction partners in vitro. For each of the AXIN1-binding partners tested, i.e. casein kinase 1 E (CK1E); c-Myc; peptidyl-prolyl cis/trans isomerase, NIMA-interacting 1 (Pin1); and p53, we found at least three different epitopes, predominantly in the central IDR of AXIN1. We functionally validated the specific AXIN1-CK1E interaction identified here with epitope-mimicking peptides and with AXIN1 variants having deletions of short binding epitopes. On the basis of these results, we propose a model in which AXIN1 competes with dishevelled (DVL) for CK1E and regulates CK1E-induced phosphorylation of DVL and activation of Wnt/-catenin signaling.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.

More info