A cell-penetrant lactam-stapled peptide for targeting eIF4E protein-protein interactions

Proteomic Characterization of Protein-Protein Interactions

Austin Biology ◽

10.26420/austinbiol.2021.1027 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Veenstra TD ◽

Keyword(s):

Protein Interactions ◽

Disease Diagnosis ◽

Cell System ◽

Protein Protein Interactions ◽

Novel Technologies ◽

Cell Functions ◽

Single Protein ◽

A Cell ◽

Molecular Components

Identifying all the molecular components within a living cell is the first step into understanding how it functions. To further understand how a cell functions requires identifying the interactions that occur between these components. This fact is especially relevant for proteins. No protein within a human cell functions on its own without interacting with another biomolecule - usually another protein. While Protein-Protein Interactions (PPI) have historically been determined by examining a single protein per study, novel technologies developed over the past couple of decades are enabling high-throughput methods that aim to describe entire protein networks within cells. In this review, some of the technologies that have led to these developments are described along with applications of these techniques. Ultimately the goal of these technologies is to map out the entire circuitry of PPI within human cells to be able to predict the global consequences of perturbations to the cell system. This predictive capability will have major impacts on the future of both disease diagnosis and treatment.

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Mitochondria: Regulators of Cell Death and Survival

The Scientific World JOURNAL ◽

10.1100/tsw.2002.809 ◽

2002 ◽

Vol 2 ◽

pp. 1569-1578 ◽

Cited By ~ 34

Author(s):

David J. Granville ◽

Roberta A. Gottlieb

Keyword(s):

Cell Death ◽

Conformational Changes ◽

Protein Interactions ◽

Apoptotic Cell ◽

Critical Role ◽

Death Process ◽

Protein Protein Interactions ◽

Ionic Changes ◽

A Cell ◽

Cyt C

The past 5 years has seen an intense surge in research devoted toward understanding the critical role of mitochondria in the regulation of cell death. Apoptosis can be initiated by a wide array of stimuli, inducing multiple signaling pathways that, for the most part, converge at the mitochondrion. Although classically considered the powerhouses of the cell, it is now understood that mitochondria are also “gatekeepers” that ultimately determine the fate of the cell. The mitochondrial decision as to whether a cell lives or dies is complex, involving protein-protein interactions, ionic changes, reactive oxygen species, and other mechanisms that require further elucidation. Once the death process is initiated, mitochondria undergo conformational changes, resulting in the release of cytochrome c (cyt c), caspases, endonucleases, and other factors leading to the onset and execution of apoptosis. The present review attempts to outline the complex milieu of events regulating the mitochondrial commitment to and processes involved in the implementation of the executioner phase of apoptotic cell death.

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Protein–protein interactions in intracellular Ca2+-release channel function

Biochemical Journal ◽

10.1042/bj3370345 ◽

1999 ◽

Vol 337 (3) ◽

pp. 345-361 ◽

Cited By ~ 8

Author(s):

John J. MACKRILL

Keyword(s):

Protein Interactions ◽

Ryanodine Receptors ◽

Accessory Proteins ◽

Protein Protein Interactions ◽

Release Channel ◽

Specific Effects ◽

Complex Protein ◽

A Cell ◽

And Function ◽

Opposing Effects

Release of Ca2+ ions from intracellular stores can occur via two classes of Ca2+-release channel (CRC) protein, the inositol 1,4,5-trisphosphate receptors (InsP3Rs) and the ryanodine receptors (RyRs). Multiple isoforms and subtypes of each CRC class display distinct but overlapping distributions within mammalian tissues. InsP3Rs and RyRs interact with a plethora of accessory proteins which modulate the activity of their intrinsic channels. Although many aspects of CRC structure and function have been reviewed in recent years, the properties of proteins with which they interact has not been comprehensively surveyed, despite extensive current research on the roles of these modulators. The aim of this article is to review the regulation of CRC activity by accessory proteins and, wherever possible, to outline the structural details of such interactions. The CRCs are large transmembrane proteins, with the bulk of their structure located cytoplasmically. Intra- and inter-complex protein–protein interactions between these cytoplasmic domains also regulate CRC function. Some accessory proteins modulate channel activity of all CRC subtypes characterized, whereas other have class- or even isoform-specific effects. Certain accessory proteins exert both direct and indirect forms of regulation on CRCs, occasionally with opposing effects. Others are themselves modulated by changes in Ca2+ concentration, thereby participating in feedback mechanisms acting on InsP3R and RyR activity. CRCs are therefore capable of integrating numerous signalling events within a cell by virtue of such protein–protein interactions. Consequently, the functional properties of InsP3Rs and RyRs within particular cells and subcellular domains are ‘customized ’ by the accessory proteins present.

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A protein tertiary structure mimetic modulator of the Hippo signalling pathway

Nature Communications ◽

10.1038/s41467-020-19224-8 ◽

2020 ◽

Vol 11 (1) ◽

Cited By ~ 1

Author(s):

Hélène Adihou ◽

Ranganath Gopalakrishnan ◽

Tim Förster ◽

Stéphanie M. Guéret ◽

Raphael Gasper ◽

...

Keyword(s):

Transcription Factor ◽

Protein Interactions ◽

Tertiary Structure ◽

Hippo Pathway ◽

Signalling Pathway ◽

Protein Protein Interactions ◽

Protein Tertiary Structure ◽

A Cell ◽

Chemical Modulation ◽

Hippo Signalling

Abstract Transcription factors are key protein effectors in the regulation of gene transcription, and in many cases their activity is regulated via a complex network of protein–protein interactions (PPI). The chemical modulation of transcription factor activity is a long-standing goal in drug discovery but hampered by the difficulties associated with the targeting of PPIs, in particular when extended and flat protein interfaces are involved. Peptidomimetics have been applied to inhibit PPIs, however with variable success, as for certain interfaces the mimicry of a single secondary structure element is insufficient to obtain high binding affinities. Here, we describe the design and characterization of a stabilized protein tertiary structure that acts as an inhibitor of the interaction between the transcription factor TEAD and its co-repressor VGL4, both playing a central role in the Hippo signalling pathway. Modification of the inhibitor with a cell-penetrating entity yielded a cell-permeable proteomimetic that activates cell proliferation via regulation of the Hippo pathway, highlighting the potential of protein tertiary structure mimetics as an emerging class of PPI modulators.

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The Hydrocarbon Staple & Beyond: Recent Advances Towards Stapled Peptide Therapeutics that Target Protein-Protein Interactions

Current Topics in Medicinal Chemistry ◽

10.2174/1568026618666180518095255 ◽

2018 ◽

Vol 18 (7) ◽

pp. 611-624 ◽

Cited By ~ 6

Author(s):

Robert A. Hillman ◽

Jonathan W. Nadraws ◽

Michael A. Bertucci

Keyword(s):

Protein Interactions ◽

Neurological Disorders ◽

Endocrine Disorders ◽

Protein Protein Interactions ◽

Peptide Therapeutics ◽

Stapled Peptide ◽

Disease States ◽

Structural Rigidity ◽

Recent Advances ◽

Pharmacokinetic Properties

Anomalous protein-protein interactions (PPIs) have been correlated to a variety of disease states, such as cancer, infectious disease, neurological disorders, diabetes, endocrine disorders and cardiovascular disease. Stapled peptides are an emerging intervention for these PPIs due to their improved structural rigidity and pharmacokinetic properties relative to unstapled peptides. This review details the most recent advances in the field of stapled peptide therapeutics, including the increasing variety of PPIs being targeted and types of peptide staples being employed.

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Grafting Hydrophobic Amino Acids Critical for Inhibition of Protein–Protein Interactions on a Cell-Penetrating Peptide Scaffold

Molecular Pharmaceutics ◽

10.1021/acs.molpharmaceut.1c00671 ◽

2021 ◽

Author(s):

Yuki Nagano ◽

Jan Vincent V. Arafiles ◽

Keiko Kuwata ◽

Yoshimasa Kawaguchi ◽

Miki Imanishi ◽

...

Keyword(s):

Amino Acids ◽

Protein Interactions ◽

Cell Penetrating Peptide ◽

Protein Protein Interactions ◽

Cell Penetrating ◽

A Cell ◽

Hydrophobic Amino Acids

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The metabolite α-KG induces GSDMC-dependent pyroptosis through death receptor 6-activated caspase-8

Cell Research ◽

10.1038/s41422-021-00506-9 ◽

2021 ◽

Author(s):

Jia-yuan Zhang ◽

Bo Zhou ◽

Ru-yue Sun ◽

Yuan-li Ai ◽

Kang Cheng ◽

...

Keyword(s):

Protein Interactions ◽

Tumor Therapy ◽

Death Receptor ◽

Caspase 8 ◽

Acidic Environment ◽

Protein Protein Interactions ◽

Regulated Cell Death ◽

A Cell ◽

Tumor Growth And Metastasis ◽

Death Receptor 6

AbstractPyroptosis is a form of regulated cell death mediated by gasdermin family members, among which the function of GSDMC has not been clearly described. Herein, we demonstrate that the metabolite α-ketoglutarate (α-KG) induces pyroptosis through caspase-8-mediated cleavage of GSDMC. Treatment with DM-αKG, a cell-permeable derivative of α-KG, elevates ROS levels, which leads to oxidation of the plasma membrane-localized death receptor DR6. Oxidation of DR6 triggers its endocytosis, and then recruits both pro-caspase-8 and GSDMC to a DR6 receptosome through protein-protein interactions. The DR6 receptosome herein provides a platform for the cleavage of GSDMC by active caspase-8, thereby leading to pyroptosis. Moreover, this α-KG-induced pyroptosis could inhibit tumor growth and metastasis in mouse models. Interestingly, the efficiency of α-KG in inducing pyroptosis relies on an acidic environment in which α-KG is reduced by MDH1 and converted to L-2HG that further boosts ROS levels. Treatment with lactic acid, the end product of glycolysis, builds an improved acidic environment to facilitate more production of L-2HG, which makes the originally pyroptosis-resistant cancer cells more susceptible to α-KG-induced pyroptosis. This study not only illustrates a pyroptotic pathway linked with metabolites but also identifies an unreported principal axis extending from ROS-initiated DR6 endocytosis to caspase-8-mediated cleavage of GSDMC for potential clinical application in tumor therapy.

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A cell-based screening method using an intracellular antibody for discovering small molecules targeting the translocation protein LMO2

Science Advances ◽

10.1126/sciadv.abg1950 ◽

2021 ◽

Vol 7 (15) ◽

pp. eabg1950

Author(s):

Nicolas Bery ◽

Carole J.R. Bataille ◽

Angela Russell ◽

Angela Hayes ◽

Florence Raynaud ◽

...

Keyword(s):

Small Molecules ◽

Protein Interactions ◽

Binding Sites ◽

Screening Method ◽

Chromosomal Translocation ◽

Resonance Method ◽

Protein Protein Interactions ◽

Intracellular Antibody ◽

A Cell

Intracellular antibodies are tools that can be used directly for target validation by interfering with properties like protein-protein interactions. An alternative use of intracellular antibodies in drug discovery is developing small-molecule surrogates using antibody-derived (Abd) technology. We previously used this strategy with an in vitro competitive surface plasmon resonance method that relied on high-affinity antibody fragments to obtain RAS-binding compounds. We now describe a novel implementation of the Abd method with a cell-based intracellular antibody–guided screening method that we have applied to the chromosomal translocation protein LMO2. We have identified a chemical series of anti-LMO2 Abd compounds that bind at the same LMO2 location as the inhibitory anti-LMO2 intracellular antibody combining site. Intracellular antibodies could therefore be used in cell-based screens to identify chemical surrogates of their binding sites and potentially be applied to any challenging proteins, such as transcription factors that have been considered undruggable.

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Purification, characterization, and cDNA cloning of an AU-rich element RNA-binding protein, AUF1

Molecular and Cellular Biology ◽

10.1128/mcb.13.12.7652-7665.1993 ◽

1993 ◽

Vol 13 (12) ◽

pp. 7652-7665 ◽

Cited By ~ 2

Author(s):

W Zhang ◽

B J Wagner ◽

K Ehrenman ◽

A W Schaefer ◽

C T DeMaria ◽

...

Keyword(s):

Protein Interactions ◽

Rna Binding ◽

K562 Cells ◽

Mrna Degradation ◽

Protein Protein Interactions ◽

Rna Recognition Motifs ◽

Reading Frame ◽

A Cell ◽

Recognition Motifs ◽

Macrophage Colony

The degradation of some proto-oncogene and lymphokine mRNAs is controlled in part by an AU-rich element (ARE) in the 3' untranslated region. It was shown previously (G. Brewer, Mol. Cell. Biol. 11:2460-2466, 1991) that two polypeptides (37 and 40 kDa) copurified with fractions of a 130,000 x g postribosomal supernatant (S130) from K562 cells that selectively accelerated degradation of c-myc mRNA in a cell-free decay system. These polypeptides bound specifically to the c-myc and granulocyte-macrophage colony-stimulating factor 3' UTRs, suggesting they are in part responsible for selective mRNA degradation. In the present work, we have purified the RNA-binding component of this mRNA degradation activity, which we refer to as AUF1. Using antisera specific for these polypeptides, we demonstrate that the 37- and 40-kDa polypeptides are immunologically cross-reactive and that both polypeptides are phosphorylated and can be found in a complex(s) with other polypeptides. Immunologically related polypeptides are found in both the nucleus and the cytoplasm. The antibodies were also used to clone a cDNA for the 37-kDa polypeptide. This cDNA contains an open reading frame predicted to produce a protein with several features, including two RNA recognition motifs and domains that potentially mediate protein-protein interactions. These results provide further support for a role of this protein in mediating ARE-directed mRNA degradation.

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