Exploration of the HIF-1α/p300 interface using peptide and Adhiron phage display technologies

Kyle, Hannah F.; Wickson, Kate; Stott, Jonathan; Burslem, George M.; Breeze, Alexander L.; Tiede, Christian; Tomlinson, Darren C.; Warriner, S. L.; Nelson, Adam; Wilson, Andrew J.; Edwards, Thomas A.

doi:10.1039/c5mb00284b

Cited by 37 publications

(55 citation statements)

References 78 publications

(213 reference statements)

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“…Peptide phage display is an increasingly common strategy to characterize known PPIs, discover new PPIs, and identify candidate peptides for PPI inhibition [16]. For example, a random-sequence peptide display library was used to characterize the interface and binding modes of the interaction between p300 and HIF-1α, and several of the identified peptides inhibited the interaction between p300 and HIF-1α with an IC 50 of <5 µM [17]. In another study, peptide phage display was used to identify the PY binding motif (LPxY or PPxY) as a strong interactor with the oxidoreductase WWOX; the authors then validated several PY-containing proteins as novel binding partners of WWOX, including the E3 ubiquitin ligase ITCH [18].…”

Section: Peptides Derived Through High-throughput Methodsmentioning

confidence: 99%

Peptides and peptidomimetics as regulators of protein–protein interactions

Cunningham

Qvit

Mochly‐Rosen

2017

Current Opinion in Structural Biology

125

107

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Protein-protein interactions are essential for almost all intracellular and extracellular biological processes. Regulation of protein-protein interactions is one strategy to regulate cell fate in a highly selective manner. Specifically, peptides are ideal candidates for inhibition of protein-protein interactions because they can mimic a protein surface to effectively compete for binding. Additionally, peptides are synthetically accessible and can be stabilized by chemical modifications. In this review, we survey screening and rational design methods for identifying peptides to inhibit protein-protein interactions, as well as methods for stabilizing peptides to effectively mimic protein surfaces. In addition, we discuss recent applications of peptides to regulate protein-protein interactions for both basic research and therapeutic purposes.

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Section: Peptides Derived Through High-throughput Methodsmentioning

confidence: 99%

Peptides and peptidomimetics as regulators of protein–protein interactions

Cunningham

Qvit

Mochly‐Rosen

2017

Current Opinion in Structural Biology

125

107

View full text Add to dashboard Cite

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“…We have previously demonstrated use of Affimers in a number of assays including immune-like (affinity) assays, in biosensors and have tested their ability to be expressed in mammalian cells to manipulate cell signalling (Tiede et al, 2014; Kyle et al, 2015; Rawlings et al, 2015; Stadler et al, 2014; Sharma et al, 2016). Here we have screened our established Affimer phage library (Tiede et al, 2014) against a broad range of targets, including homologous protein family members, to isolate highly specific and renewable binding reagents that can be used both in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

Affimer proteins are versatile and renewable affinity reagents

Tiede

Bedford

Heseltine

et al. 2017

eLife

Self Cite

160

162

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Molecular recognition reagents are key tools for understanding biological processes and are used universally by scientists to study protein expression, localisation and interactions. Antibodies remain the most widely used of such reagents and many show excellent performance, although some are poorly characterised or have stability or batch variability issues, supporting the use of alternative binding proteins as complementary reagents for many applications. Here we report on the use of Affimer proteins as research reagents. We selected 12 diverse molecular targets for Affimer selection to exemplify their use in common molecular and cellular applications including the (a) selection against various target molecules; (b) modulation of protein function in vitro and in vivo; (c) labelling of tumour antigens in mouse models; and (d) use in affinity fluorescence and super-resolution microscopy. This work shows that Affimer proteins, as is the case for other alternative binding scaffolds, represent complementary affinity reagents to antibodies for various molecular and cell biology applications.DOI: http://dx.doi.org/10.7554/eLife.24903.001

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“…For example, peptidomimetics based on Helix 2 mimic the orientation of residues 815–823, including two conserved leucine residues (L818 and L822 in HIF‐1α) that bind in a hydrophobic pocket in CBP/p300 (Figure b). Phage display‐based analyses suggest that Helix 2 binds p300 with higher affinity than any other region of the HIF‐1α CAD, which could explain why most of the reported peptidomimetic compounds have targeted this helix.…”

Section: Inhibition Of Protein–protein Interactions In the Hif Systemmentioning

confidence: 99%

Targeting Protein–Protein Interactions in the HIF System

et al. 2016

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Animals respond to chronic hypoxia by increasing the levels of a transcription factor known as the hypoxia‐inducible factor (HIF). HIF upregulates multiple genes, the products of which work to ameliorate the effects of limited oxygen at cellular and systemic levels. Hypoxia sensing by the HIF system involves hydroxylase‐catalysed post‐translational modifications of the HIF α‐subunits, which 1) signal for degradation of HIF‐α and 2) limit binding of HIF to transcriptional coactivator proteins. Because the hypoxic response is relevant to multiple disease states, therapeutic manipulation of the HIF‐mediated response has considerable medicinal potential. In addition to modulation of catalysis by the HIF hydroxylases, the HIF system manifests other possibilities for therapeutic intervention involving protein–protein and protein–nucleic acid interactions. Recent advances in our understanding of the structural biology and biochemistry of the HIF system are facilitating medicinal chemistry efforts. Herein we give an overview of the HIF system, focusing on structural knowledge of protein–protein interactions and how this might be used to modulate the hypoxic response for therapeutic benefit.

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Exploration of the HIF-1α/p300 interface using peptide and Adhiron phage display technologies

Cited by 37 publications

References 78 publications

Peptides and peptidomimetics as regulators of protein–protein interactions

Peptides and peptidomimetics as regulators of protein–protein interactions

Affimer proteins are versatile and renewable affinity reagents

Targeting Protein–Protein Interactions in the HIF System

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