Abstract
Metazoan proteomes contain many paralogous proteins that have evolved distinct functions. The Ena/VASP family of actin regulators consists of three members that share an EVH1 interaction domain with a 100% conserved binding site. A proteome-wide screen revealed photoreceptor cilium actin regulator (PCARE) as a high-affinity ligand for ENAH EVH1. Here, we report the surprising observation that PCARE is ~100-fold specific for ENAH over paralogs VASP and EVL and can selectively bind ENAH and inhibit ENAH-dependent adhesion in cells. Specificity arises from a mechanism whereby PCARE stabilizes a conformation of the ENAH EVH1 domain that is inaccessible to family members VASP and EVL. Structure-based modeling rapidly identified seven residues distributed throughout EVL that are sufficient to differentiate binding by ENAH vs. EVL. By exploiting the ENAH-specific conformation, we rationally designed the tightest and most selective ENAH binder to date. Our work uncovers a conformational mechanism of interaction specificity that distinguishes highly similar paralogs and establishes tools for dissecting specific Ena/VASP functions in processes including cancer cell invasion.
Original language | English (US) |
---|---|
Article number | e70601 |
Journal | eLife |
Volume | 10 |
DOIs | |
State | Published - Dec 2021 |
ASJC Scopus subject areas
- General Neuroscience
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology
Fingerprint
Dive into the research topics of 'A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers'. Together they form a unique fingerprint.Datasets
-
ENAH EVH1 domain bound to peptide from protein PCARE
Hwang, T. (Contributor), Parker, S. S. (Contributor), Hill, S. M. (Contributor), Ilunga, M. W. (Contributor), Grant, R. A. (Contributor), Mouneimne, G. (Contributor) & Keating, A. E. (Contributor), Protein Data Bank (PDB), Nov 24 2021
DOI: 10.2210/pdb7LXF/pdb, https://www.wwpdb.org/pdb?id=pdb_00007lxf
Dataset