TY  - JOUR
A1  - Sundell, Gustav N.
A1  - Arnold, Roland
A1  - Ali, Muhammad
A1  - Naksukpaiboon, Piangfan
A1  - Orts, Julien
A1  - Güntert, Peter
A1  - Chi, Celestine N.
A1  - Ivarsson, Ylva
T1  - Proteome‐wide analysis of phospho‐regulated PDZ domain interactions
T2  - Molecular systems biology
N2  - A key function of reversible protein phosphorylation is to regulate protein–protein interactions, many of which involve short linear motifs (3–12 amino acids). Motif‐based interactions are difficult to capture because of their often low‐to‐moderate affinities. Here, we describe phosphomimetic proteomic peptide‐phage display, a powerful method for simultaneously finding motif‐based interaction and pinpointing phosphorylation switches. We computationally designed an oligonucleotide library encoding human C‐terminal peptides containing known or predicted Ser/Thr phosphosites and phosphomimetic variants thereof. We incorporated these oligonucleotides into a phage library and screened the PDZ (PSD‐95/Dlg/ZO‐1) domains of Scribble and DLG1 for interactions potentially enabled or disabled by ligand phosphorylation. We identified known and novel binders and characterized selected interactions through microscale thermophoresis, isothermal titration calorimetry, and NMR. We uncover site‐specific phospho‐regulation of PDZ domain interactions, provide a structural framework for how PDZ domains accomplish phosphopeptide binding, and discuss ligand phosphorylation as a switching mechanism of PDZ domain interactions. The approach is readily scalable and can be used to explore the potential phospho‐regulation of motif‐based interactions on a large scale.
KW  - PDZ domain
KW  - phage display
KW  - phosphorylation
KW  - protein–protein interaction
KW  - Scribble
Y1  - 2018
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/47144
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-471443
SN  - 1744-4292
N1  - This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
VL  - 14
IS  - 8, e8129
SP  - 1
EP  - 22
PB  - EMBO Press
CY  - Heidelberg
ER  -