{"doi":"10.7554/elife.70680","title":"Native proline-rich motifs exploit sequence context to target actin-remodeling Ena/VASP protein ENAH","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.","journal":"eLife","year":2022,"id":249647,"datarank":0.9663735055029499,"base_score":3.332204510175204,"endowment":3.332204510175204,"self_citation_contribution":0.49983067652628066,"citation_network_contribution":0.46654282897666915,"self_endowment_contribution":0.49983067652628066,"citer_contribution":0.46654282897666915,"corpus_percentile":null,"corpus_rank":null,"citation_count":27,"citer_count":27,"citers_with_citation_signal":22,"citers_with_endowment":22,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9556,"is_data_producer":true,"deposit_databanks":{"GEO":["GSE166938"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":754634,"name":"Sara S Parker","orcid":"0000-0003-3670-6147","position":1,"is_corresponding":false},{"id":754635,"name":"Samantha M Hill","orcid":"0000-0002-6454-7430","position":2,"is_corresponding":false},{"id":281464,"name":"Robert A. Grant","orcid":"0000-0002-5072-2867","position":3,"is_corresponding":false},{"id":755332,"name":"Meucci W Ilunga","orcid":null,"position":4,"is_corresponding":false},{"id":825105,"name":"Venkatesh Sivaraman","orcid":"0000-0002-6965-3961","position":5,"is_corresponding":false},{"id":754636,"name":"Ghassan Mouneimne","orcid":"0000-0001-8103-4701","position":6,"is_corresponding":false},{"id":261779,"name":"Amy E. Keating","orcid":"0000-0003-4074-8980","position":7,"is_corresponding":false},{"id":754633,"name":"Theresa Hwang","orcid":"0000-0002-9011-2174","position":0,"is_corresponding":true}],"reference_count":66,"raw_metadata":null,"created_at":"2026-07-19T00:24:23.754337Z","pmid":"35076015","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}