A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers

Theresa Hwang; Sara S. Parker; Samantha M. Hill; Meucci W. Ilunga; Robert A. Grant; Ghassan Mouneimne; Amy E. Keating

doi:10.7554/eLife.70601

A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers

Theresa Hwang, Sara S. Parker, Samantha M. Hill, Meucci W. Ilunga, Robert A. Grant, Ghassan Mouneimne, Amy E. Keating

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

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	https://doi.org/10.7554/eLife.70601
State	Published - Dec 2021
Externally published	Yes

ASJC Scopus subject areas

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.7554/eLife.70601

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@article{6efcde3f041d4a81a59950e12ce723c8,

title = "A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers",

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.",

author = "Theresa Hwang and Parker, {Sara S.} and Hill, {Samantha M.} and Ilunga, {Meucci W.} and Grant, {Robert A.} and Ghassan Mouneimne and Keating, {Amy E.}",

note = "Funding Information: This project was supported by NIGMS award R01 GM129007 to AEK and by NCI award R01 CA196885-01 to GM. Part of this work has been supported by Koch Institute NCI Cancer Center (Core) Support Grant (CCSG) P30-CA14051. Part of this work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P30 GM124165). The Eiger 16 M detector on the 24-ID-E beamline is funded by an NIH-ORIP HEI grant (S10OD021527). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract DE-AC02-06CH11357. TH was partially supported by NIGMS T32 GM007287 and a fellowship from the Koch Institute for Integrative Cancer Research. Publisher Copyright: {\textcopyright} Hwang et al.",

year = "2021",

month = dec,

doi = "10.7554/eLife.70601",

language = "English (US)",

volume = "10",

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T1 - A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers

AU - Hwang, Theresa

AU - Parker, Sara S.

AU - Hill, Samantha M.

AU - Ilunga, Meucci W.

AU - Grant, Robert A.

AU - Mouneimne, Ghassan

AU - Keating, Amy E.

N1 - Funding Information: This project was supported by NIGMS award R01 GM129007 to AEK and by NCI award R01 CA196885-01 to GM. Part of this work has been supported by Koch Institute NCI Cancer Center (Core) Support Grant (CCSG) P30-CA14051. Part of this work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P30 GM124165). The Eiger 16 M detector on the 24-ID-E beamline is funded by an NIH-ORIP HEI grant (S10OD021527). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract DE-AC02-06CH11357. TH was partially supported by NIGMS T32 GM007287 and a fellowship from the Koch Institute for Integrative Cancer Research. Publisher Copyright: © Hwang et al.

PY - 2021/12

Y1 - 2021/12

N2 - 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.

AB - 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.

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SN - 2050-084X

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A distributed residue network permits conformational binding specificity in a conserved family of actin remodelers

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