PT  - JOURNAL ARTICLE
AU  - Pau Jané
AU  - Gergő Gógl
AU  - Camille Kostmann
AU  - Goran Bich
AU  - Virginie Girault
AU  - Célia Caillet-Saguy
AU  - Pascal Eberling
AU  - Renaud Vincentelli
AU  - Nicolas Wolff
AU  - Gilles Travé
AU  - Yves Nominé
TI  - Interactomic affinity profiling by holdup assay: acetylation and distal residues impact the PDZome-binding specificity of PTEN phosphatase
AID  - 10.1101/2020.07.01.181487
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.07.01.181487
4099  - http://biorxiv.org/content/early/2020/07/02/2020.07.01.181487.short
4100  - http://biorxiv.org/content/early/2020/07/02/2020.07.01.181487.full
AB  - Protein domains often recognize short linear protein motifs composed of a core conserved consensus sequence surrounded by less critical, modulatory positions. Here we used an accurate experimental approach combining high-throughput holdup chromatographic assay and fluorescence polarization to measure quantitative binding affinity profiles of the PDZ domain-binding motif (PBM) of PTEN phosphatase towards the 266 known human PDZ domains. Inclusion of N-terminal flanking residues, acetylation or mutation of a lysine at a modulatory position significantly altered the PDZome-binding profile of the PTEN PBM. A specificity index is also introduced to quantify the specificity of a given PBM over the complete PDZome. Our results highlight the impact of modulatory residues and post-translational modifications on PBM interactomes and their specificity.Competing Interest StatementThe authors have declared no competing interest.