PT  - JOURNAL ARTICLE
AU  - Theresa Hwang
AU  - Robert A. Grant
AU  - Meucci W. Ilunga
AU  - Venkatesh Sivaraman
AU  - Amy E. Keating
TI  - Native proline-rich motifs exploit sequence context to target actin-remodeling Ena/VASP proteins
AID  - 10.1101/2021.03.22.436451
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.03.22.436451
4099  - http://biorxiv.org/content/early/2021/06/18/2021.03.22.436451.short
4100  - http://biorxiv.org/content/early/2021/06/18/2021.03.22.436451.full
AB  - The human proteome is replete with short linear motifs (SLiMs) of 4-6 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 systematically examine the influence of SLiM sequence context on protein-protein interactions. Focusing on the EVH1 domain of ENAH, an actin regulator that is upregulated in invasive cancers, we screened 36-residue proteome-derived peptides for binding. We discovered a pocket on the ENAH EVH1 domain that diverged from its orthologs to recognize extended SLiMs, and we found that proteins with two EVH1-binding SLiMs can wrap around a single domain. We also found that the ciliary protein PCARE uses an extended 23-residue region to obtain higher affinity than any known ENAH EVH1-binding motif. Our screen provides a way to uncover the effects of broader proteomic context on motif-mediated interactions, revealing diverse mechanisms of contextual control over EVH1 interactions and establishing that SLiMs can’t be fully understood outside of their native context.Competing Interest StatementThe authors have declared no competing interest.