Rational design of EGFR dimerization-disrupting peptides: A new strategy to combat drug resistance in targeted lung cancer therapy

Liu, Qiuhong; Zhou, Jingxin; Gao, Jing; Ma, Wentao; Wang, Shilei; Lü, Xing

doi:10.1016/j.biochi.2020.07.010

Cited by 5 publications

(5 citation statements)

References 44 publications

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“…In addition, the EGFR KD-C -nBS (K d = 16 μM) and EGFR KD-C -nCBS (K d = 38 μM) interactions failed to be detected. These K d values were determined by measuring the association between the recombinant protein and synthetic peptides using fluorescence anisotropy in vitro 26 , which may not reflect the binding properties in the cytoplasm of mammalian cells. Since the EGFR KD-C -Mig6seg1 (K d = 13 μM) interaction, which had been confirmed in mammalian cells in the previous literature 27 , was able to be detected, H-SOLIS may be useful as a tool for reliable detection of specific binders in the cytosol of living cells.…”

Section: Discussionmentioning

confidence: 99%

“…The C-lobe of one receptor chain (activator) docks with the N-lobe of the other (receiver), which triggers activation of the kinase domain 25 . A recent study identified two N-lobe-derived peptides named N-terminal binding sequence (nBS) and N-terminal core binding sequence (nCBS) that could bind to the C-lobe 26 . Furthermore, a peptide segment derived from mitogeninducible gene 6 (Mig6seg1) binds to the C-lobe and inhibits dimerization of EGFR 27 .…”

Section: Demonstrating Versatility Of H-solismentioning

confidence: 99%

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A growth-based platform for detecting domain–peptide interactions in the cytoplasm of mammalian cells

Kimura

Kashima

Kawahara

2022

Sci Rep

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Development of a method for detecting protein–protein interactions (PPIs) in living cells is important for therapeutic drug screening against various diseases including infectious diseases. We have recently developed a method named SOS localization-based interaction screening (SOLIS), in which we designed membrane-anchored and SOS-fused chimeric proteins, whose PPI-dependent association triggers membrane localization of the SOS-fused chimeric protein, activates the Ras/MAPK pathway, and induces cell growth. While SOLIS was able to detect relatively strong PPIs, further sensitivity was required for detecting intracellular endogenous PPIs typically having a micromolar order of dissociation constant (Kd). Here we develop high-sensitive SOLIS (H-SOLIS) that could universally detect PPIs with lower affinities. In order to improve the sensitivity, H-SOLIS introduces a heterodimeric helper interaction, in which addition of a small-molecule helper ligand could accommodate association of the two chimeric proteins and regulate the sensitivity. Four types of domain–peptide interactions having known Kd values are employed to examine the versatility and detection limit of H-SOLIS. Consequently, the heterodimer-inducible helper ligand dramatically enhances detection sensitivity, lowering the detection limit to a ten-micromolar order of Kd. Thus, H-SOLIS could be a platform to detect disease-related domain–peptide interactions for drug discovery screening.

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Section: Discussionmentioning

confidence: 99%

Section: Demonstrating Versatility Of H-solismentioning

confidence: 99%

A growth-based platform for detecting domain–peptide interactions in the cytoplasm of mammalian cells

Kimura

Kashima

Kawahara

2022

Sci Rep

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“…60 Peptides that disrupt EGFR dimerization have recently been investigated as a potential treatment option for drug-resistant cancers, suggesting that disrupting EGFR dimerization may be a viable approach for future drug development. 61,62 SAXS is well suited to play an important role as a high-throughput screening method for EGFR dimerization, providing a direct structural readout of a drug candidate's ability to impact dimer formation. To demonstrate this, the theoretical X-ray scattering profiles of the EGFR monomer and dimer (PDB ID: 5WB7 chains B and C) were determined using the Fast Xray Scattering (FoXS) web server (Figure 3A).…”

Section: ■ Cryogenic Electron Microscopy (Cryoem)mentioning

confidence: 99%

“…Using SAXS, Hu et al investigated the effect of extracellular mutations on the structure of EGFR dimers and found that they resulted in enhanced dimerization upon binding of ligands that normally induce weaker dimers, suggesting that these mutations impair ligand discrimination and increase the amount of active EGFR dimers . Peptides that disrupt EGFR dimerization have recently been investigated as a potential treatment option for drug-resistant cancers, suggesting that disrupting EGFR dimerization may be a viable approach for future drug development. , SAXS is well suited to play an important role as a high-throughput screening method for EGFR dimerization, providing a direct structural readout of a drug candidate’s ability to impact dimer formation. To demonstrate this, the theoretical X-ray scattering profiles of the EGFR monomer and dimer (PDB ID: 5WB7 chains B and C) were determined using the Fast X-ray Scattering (FoXS) web server (Figure A).…”

Section: Small Angle X-ray Scattering (Saxs)mentioning

confidence: 99%

Evolving Experimental Techniques for Structure-Based Drug Design

et al. 2022

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Structure-based drug design (SBDD) is a prominent method in rational drug development and has traditionally benefitted from the atomic models of protein targets obtained using X-ray crystallography at cryogenic temperatures. In this perspective, we highlight recent advances in the development of structural techniques that are capable of probing dynamic information about protein targets. First, we discuss advances in the field of X-ray crystallography including serial room-temperature crystallography as a method for obtaining high-resolution conformational dynamics of protein-inhibitor complexes. Next, we look at cryogenic electron microscopy (cryoEM), another high-resolution technique that has recently been used to study proteins and protein complexes that are too difficult to crystallize. Finally, we present small-angle X-ray scattering (SAXS) as a potential high-throughput screening tool to identify inhibitors that target protein complexes and protein oligomerization.

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“…Small molecule allosteric inhibitors such as EAI045 and JBJ-04-125-02 inhibit the kinase activity of oncogenic EGFR variants (most notably L858R/T790M/C797S EGFR) by binding to an allosteric pocket located within the kinase domain. In addition, there are now several reports − of peptide or peptidomimetic inhibitors that target the JM segment, an intracellular region whose dimerization is essential for kinase activation. − One set of inhibitors consists of cell-permeant hydrocarbon-stapled peptides − designed to inhibit EGFR by blocking the formation of the antiparallel JM coiled coil dimer within the receptor cytoslic domain (Figure ). , …”

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confidence: 99%

Allosteric Inhibition of the Epidermal Growth Factor Receptor

et al. 2021

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We previously reported a family of hydrocarbon-stapled peptides designed to interact with the epidermal growth factor receptor (EGFR) juxtamembrane (JM) segment, blocking its ability to form a coiled coil dimer that is essential for receptor activation. These hydrocarbon-stapled peptides, most notably E1S, decreased the proliferation of cell lines that express wild-type EGFR (H2030 and A431) as well as those expressing the oncogenic mutants EGFR L858R (H3255) and L858R/T790M (H1975). Although our previous investigations provided evidence that E1S interacted with EGFR directly, the location and details of these interactions were not established. Here we apply biochemical and cross-linking mass spectrometry tools to better define the interactions between E1S and EGFR. Taken with previously reported structure–activity relationships, our results support a model in which E1S interacts simultaneously with both the JM and the C-lobe of the activator kinase, effectively displacing the JM of the receiver kinase. Our results also reveal potential interactions between E1S and the N-terminal region of the C-terminal tail. We propose a model in which E1S inhibits EGFR by both mimicking and inhibiting JM coiled coil formation. This model could be used to design novel, allosteric (and perhaps nonpeptidic) EGFR inhibitors.

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Rational design of EGFR dimerization-disrupting peptides: A new strategy to combat drug resistance in targeted lung cancer therapy

Cited by 5 publications

References 44 publications

A growth-based platform for detecting domain–peptide interactions in the cytoplasm of mammalian cells

A growth-based platform for detecting domain–peptide interactions in the cytoplasm of mammalian cells

Evolving Experimental Techniques for Structure-Based Drug Design

Allosteric Inhibition of the Epidermal Growth Factor Receptor

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