A modular toolkit to inhibit proline-rich motif–mediated protein–protein interactions

Abstract

Small-molecule competitors of protein-protein interactions are urgently needed for functional analysis of large-scale genomics and proteomics data. Particularly abundant, yet so far undruggable, targets include domains specialized in recognizing proline-rich segments, including Src-homology 3 (SH3), WW, GYF, and Drosophila enabled (Ena)/vasodilator-stimulated phosphoprotein (VASP) homology 1 (EVH1) domains. Here, we present a modular strategy to obtain an extendable toolkit of chemical fragments (ProMs) designed to replace pairs of conserved prolines in recognition motifs. As proofof-principle, we developed a small, selective, peptidomimetic inhibitor of Ena/VASP EVH1 domain interactions. Highly invasive MDA MB 231 breast-cancer cells treated with this ligand showed displacement of VASP from focal adhesions, as well as from the front of lamellipodia, and strongly reduced cell invasion. General applicability of our strategy is illustrated by the design of an ErbB4-derived ligand containing two ProM-1 fragments, targeting the yes-associated protein 1 (YAP1)-WW domain with a fivefold higher affinity.Ena | VASP | protein-protein interaction | actin cytoskeleton | cell migration

Structural enzymology binding studies of the peptide‐substrate‐binding domain of human collagen prolyl 4‐hydroxylase (type‐II): High affinity peptides have a PxGP sequence motif

“…Proline‐rich peptide segments (PRSs) are commonly found at the interfaces of protein–protein interaction sites . The protein domains that bind such PRSs, referred to as PRS‐recognizing domains (PRDs) are involved in a wide range of functions.…”

Network analysis of mitonuclear GWAS reveals functional networks and tissue expression profiles of disease-associated genes

“…STK35/Clik1 regulates gene expression, while the PDZ-LIM family comprises stress fiber-associated proteins that control regulatory factors through sequestration in the cytoplasm; at least one PDLIM family member has been shown to directly target STK35 to stress fibers (Vallenius and Makela 2002). The other gene products in the cytoskeletal cluster, CFL1 (cofilin 1), ACTN1 (alpha actinin 1), and ERBB4 (encoding receptor tyrosine protein kinase erbB-4), all interact to regulate actin stress fibers (Hirata et al 2015; Opitz et al 2015). Moreover, the STK35 homologue STK35L1 is a known regulator of the tumor suppressor p16(INK4a) (Goyal et al 2011).…”

“…The cancer intrinsic apoptosis eQTL findings demonstrate that the directional impact of risk variants on gene expression can predict target pathways for intervention. Pharmaceutical agents targeting the intrinsic apoptotic pathway are already in experimental and clinical use (Almstedt and Schmidt 2015; Mohana-Kumaran et al 2014; Mukherjee et al 2015; Opitz et al 2015; Paraiso et al 2012; Subramaniam et al 2015; Yaswen et al 2015), so it is readily conceivable that cancer patients could be screened for variants in this hub and treatment designed based on their genotype. In addition, our findings of tissue specificity in certain disease gene sets reinforces the importance of considering tissue specificity in drug screening and development, while also suggesting that interventions targeted to the tissue of interest may provide greater benefit with fewer off-target effects.…”

Passende Kleider für Proteine schneidern

“…Die Kombination zweier unterschiedlicher Einheiten ergab ein Heterodimer, das spezifisch an die EVH1‐Domäne des Proteins EnaH bindet (Kd = 4 μM) (Abbildung 5). Hoch aggressive Brustkrebszelllinien wie MDA MB 231 wurden mit diesem Liganden behandelt, und die Zellinvasion stoppte 11…”