Large-scale phosphomimetic screening identifies phospho-modulated motif-based protein interactions.

Autor: Kliche J; Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden., Garvanska DH; Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark., Simonetti L; Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden., Badgujar D; Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden., Dobritzsch D; Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden., Nilsson J; Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark., Davey NE; Division of Cancer Biology, The Institute of Cancer Research, London, UK., Ivarsson Y; Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden.
Jazyk: angličtina
Zdroj: Molecular systems biology [Mol Syst Biol] 2023 Jul 11; Vol. 19 (7), pp. e11164. Date of Electronic Publication: 2023 May 23.
DOI: 10.15252/msb.202211164
Abstrakt: Phosphorylation is a ubiquitous post-translation modification that regulates protein function by promoting, inhibiting or modulating protein-protein interactions. Hundreds of thousands of phosphosites have been identified but the vast majority have not been functionally characterised and it remains a challenge to decipher phosphorylation events modulating interactions. We generated a phosphomimetic proteomic peptide-phage display library to screen for phosphosites that modulate short linear motif-based interactions. The peptidome covers ~13,500 phospho-serine/threonine sites found in the intrinsically disordered regions of the human proteome. Each phosphosite is represented as wild-type and phosphomimetic variant. We screened 71 protein domains to identify 248 phosphosites that modulate motif-mediated interactions. Affinity measurements confirmed the phospho-modulation of 14 out of 18 tested interactions. We performed a detailed follow-up on a phospho-dependent interaction between clathrin and the mitotic spindle protein hepatoma-upregulated protein (HURP), demonstrating the essentiality of the phospho-dependency to the mitotic function of HURP. Structural characterisation of the clathrin-HURP complex elucidated the molecular basis for the phospho-dependency. Our work showcases the power of phosphomimetic ProP-PD to discover novel phospho-modulated interactions required for cellular function.
(© 2023 The Authors. Published under the terms of the CC BY 4.0 license.)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje