High-throughput competitive fluorescence polarization assay reveals functional redundancy in the S100 protein family.
Fiche publication
Date publication
juillet 2020
Journal
The FEBS journal
Auteurs
Membres identifiés du Cancéropôle Est :
Dr GOGL Gergo
Tous les auteurs :
Simon MA, Ecsédi P, Kovács GM, Póti ÁL, Reményi A, Kardos J, Gógl G, Nyitray L
Lien Pubmed
Résumé
The calcium-binding, vertebrate-specific S100 protein family consists of 20 paralogs in humans (referred as the S100ome), with several clinically important members. To explore their protein-protein interactions (PPIs) quantitatively, we have chosen an unbiased, high-throughput, competitive fluorescence polarization (FP) assay that revealed a partial functional redundancy when the complete S100ome (n = 20) was tested against numerous model partners (n = 13). Based on their specificity, the S100ome can be grouped into two distinct classes: promiscuous and orphan. In the first group, members bound to several ligands (> 4-5) with comparable high affinity, while in the second one, the paralogs bound only one partner weakly, or no ligand was identified. Our results demonstrate that FP assays are highly suitable for quantitative interaction profiling of selected protein families. Moreover, we provide evidence that PPI-based phenotypic characterization can complement or even exceed the information obtained from the sequence-based phylogenetic analysis of the S100ome, an evolutionary young protein family.
Mots clés
calcium, fluorescence anisotropy, isothermal titration calorimetry, systems biology
Référence
FEBS J. 2020 07;287(13):2834-2846