7ZX4

Clathrin N-terminal domain in complex with a HURP phospho-peptide

Summary for 7ZX4

• Entry DOI: 10.2210/pdb7zx4/pdb
• Descriptor: Clathrin heavy chain 1, Disks large-associated protein 5, GLYCEROL, ... (6 entities in total)
• Functional Keywords: complex, phospho-regulated slim-based interactions, hurp, dlgap5, endocytosis
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 5
• Total formula weight: 88717.62

Authors

Kliche, J.,Badgujar, D.,Dobritzsch, D.,Ivarsson, Y. (deposition date: 2022-05-20, release date: 2023-03-29, Last modification date: 2024-02-07)

Primary citation

Kliche, J.,Garvanska, D.H.,Simonetti, L.,Badgujar, D.,Dobritzsch, D.,Nilsson, J.,Davey, N.E.,Ivarsson, Y.
Large-scale phosphomimetic screening identifies phospho-modulated motif-based protein interactions.
Mol.Syst.Biol., 19:e11164-e11164, 2023

PubMed Abstract: 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.

PubMed: 37219487
DOI: 10.15252/msb.202211164

Experimental method

X-RAY DIFFRACTION (2.08 Å)