Native proline-rich motifs exploit sequence context to target actin-remodeling Ena/VASP protein ENAH
Name
elife-70680-v1.pdf
Description
Published version
Size
3.56 MB
Format
Adobe PDF
Checksum (MD5)
a95985be8a3cf5181fed111d62e58273
Author(s)
Hwang, Theresa
Parker, Sara S
Hill, Samantha M
Grant, Robert A
Ilunga, Meucci W
Sivaraman, Venkatesh
Mouneimne, Ghassan
Keating, Amy E
Date Issued
2022
Journal
eLife
Publisher
eLife Sciences Publications, Ltd
Citation
Hwang, Theresa, Parker, Sara S, Hill, Samantha M, Grant, Robert A, Ilunga, Meucci W et al. 2022. "Native proline-rich motifs exploit sequence context to target actin-remodeling Ena/VASP protein ENAH." eLife, 11.
Version
Final published version
Abstract
The human proteome is replete with short linear motifs (SLiMs) of four to six residues that are critical for protein-protein interactions, yet the importance of the sequence surrounding such motifs is underexplored. We devised a proteomic screen to examine the influence of SLiM sequence context on protein-protein interactions. Focusing on the EVH1 domain of human ENAH, an actin regulator that is highly expressed in invasive cancers, we screened 36-residue proteome-derived peptides and discovered new interaction partners of ENAH and diverse mechanisms by which context influences binding. A pocket on the ENAH EVH1 domain that has diverged from other Ena/VASP paralogs recognizes extended SLiMs and favors motif-flanking proline residues. Many high-affinity ENAH binders that contain two proline-rich SLiMs use a noncanonical site on the EVH1 domain for binding and display a thermodynamic signature consistent with the two-motif chain engaging a single domain. We also found that photoreceptor cilium actin regulator (PCARE) uses an extended 23-residue region to obtain a higher affinity than any known ENAH EVH1-binding motif. Our screen provides a way to uncover the effects of proteomic context on motif-mediated binding, revealing diverse mechanisms of control over EVH1 interactions and establishing that SLiMs can’t be fully understood outside of their native context.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.7554/ELIFE.70680
