A simple method for developing lysine targeted covalent protein reagents

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Dec. 1, 2023

Abstract Peptide-based covalent probes can target shallow protein surfaces not typically addressable using small molecules, yet there is a need for versatile approaches to convert native peptide sequences into binders that broad range of residues. Here we report protein-based thio-methacrylate esters—electrophiles be installed easily on unprotected peptides and proteins via cysteine side chains, react efficiently selectively with lysine chains the target. Methacrylate phosphopeptides derived from 14-3-3-binding irreversibly label 14-3-3σ either or residues, depending position electrophile. targeting conserved residue exhibit pan-isoform binding 14-3-3 both in lysates extracellular media. Finally, apply this approach develop binders. A methacrylate-modified variant colicin E9 immunity binds DNAse, resulting significantly higher thermal stability relative non-covalent complex. Our offers simple route potent

Language: Английский

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Mitochondrial Protease Targeting Chimeras for Mitochondrial Matrix Protein Degradation

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(23), P. 12861 - 12869

Published: June 5, 2023

Targeted protein degradation (TPD) is an emerging technique for regulation. Currently, all TPD developed in eukaryotic cells relies on either ubiquitin-proteasome or lysosomal systems, thus are powerless against target proteins membrane organelles lacking proteasomes and lysosomes, such as mitochondria. Here, we a mitochondrial protease targeting chimera (MtPTAC) to address this issue. MtPTAC bifunctional small molecule that can bind caseinolytic P (ClpP) at one end the other. Mechanistically, activates hydrolase activity of ClpP while simultaneously bringing into proximity with ClpP. Taking RNA polymerase (POLRMT) model protein, have demonstrated powerful proteolytic ability antitumor application prospects MtPTAC, both vivo vitro. This first modularly designed specifically hydrolyze inside

Language: Английский

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Development and Applications of Chimera Platforms for Tyrosine Phosphorylation

ACS Central Science, Journal Year: 2023, Volume and Issue: 9(8), P. 1558 - 1566

Published: Aug. 9, 2023

Chimeric small molecules that induce post-translational modification (PTM) on a target protein by bringing it into proximity to PTM-inducing enzyme are furnishing novel modalities perturb function. Despite recent advances, such unavailable for critical PTM, tyrosine phosphorylation. Furthermore, the contemporary design paradigm of chimeric molecules, formed joining noninhibitory binder with protein, prohibits recruitment most enzymes as their binders unavailable. Here, we report two platforms generate phosphorylation-inducing (PHICS) We PHICS from both (scantily available, platform 1) and kinase inhibitors (abundantly 2) using cysteine-based group transfer chemistry. triggered phosphorylation residues in diverse sequence contexts proteins (e.g., membrane-associated, cytosolic) displayed multiple bioactivities, including initiation growth receptor signaling cascade death drug-resistant cancer cells. These studies provide an approach biologically relevant PTM lay foundation pharmacologic editing (i.e., induction or removal) abundantly available -erasing enzymes.

Language: Английский

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Ligand‐Directed Chemistry for Protein Labeling for Affinity‐Based Protein Analysis

Israel Journal of Chemistry, Journal Year: 2023, Volume and Issue: 63(3-4)

Published: Feb. 6, 2023

Abstract Structural and functional analyses of proteins‐of‐interest (POI) in multimolecular crowding conditions (mMCC) such as live cells tissues are regarded inevitable challenges for depth understanding the real shapes POIs their existing natural environments. Activity‐based protein profiling (ABPP) is a definitely powerful tool capable analyzing proteome possessing particular activity under mMCC. While ABPP usually targets interest, study mMCC also valuable. Although activity‐based probes (ABPs) often used this aim, most conventional ABPs cause loss original activities, therefore not perfectly suitable analysis labeled proteins. Ligand‐directed chemistry (LDchem) developed by our group an alternative approach ABPs, that can modify surface POI rather than its active site using cleavable electrophile traceless manner. LDchem thus enables labeling with synthetic fluorophore no or minimal effects on functions even In review, we briefly describe principle native summarize recent chemical biology applications imaging‐based biological construction POI‐based biosensors.

Language: Английский

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Covalent PROTAC design method based on a sulfonyl pyridone probe

Chemical Communications, Journal Year: 2023, Volume and Issue: 60(6), P. 686 - 689

Published: Nov. 29, 2023

Covalent proteolysis-targeting chimeras (PROTACs) offer enhanced selectivity, prolonged action, and increased efficacy against challenging target proteins.

Language: Английский

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oxSTEF Reagents Are Tunable and Versatile Electrophiles for Selective Disulfide-Rebridging of Native Proteins

Bioconjugate Chemistry, Journal Year: 2023, Volume and Issue: 34(6), P. 994 - 1003

Published: May 18, 2023

Site-selective disulfide rebridging has emerged as a powerful strategy to modulate the structural and functional properties of proteins. Here, we introduce novel class electrophilic reagents, designated oxSTEF, that demonstrate excellent efficiency in via double thiol exchange. The oxSTEF reagents are prepared using an efficient synthetic sequence which may be diverted obtain range derivatives allowing for tuning reactivity or steric bulk. We highly selective cyclic peptides native proteins, such human growth hormone, absence cross-reactivity with other nucleophilic amino acid residues. conjugates undergo glutathione-mediated disintegration under tumor-relevant glutathione concentrations, highlights their potential use targeted drug delivery. Finally, α-dicarbonyl motif enables "second phase" oxime ligation, furthermore increases stability significantly.

Language: Английский

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Proximity-Enhanced Cysteine-Histidine Crosslinking for Elucidating Intrinsically Disordered and Other Protein Complexes

Chemical Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

A novel proximity-enhanced crosslinker using histidine–cysteine trapping uncovers a cryptic 14-3-3/hyperphosphorylated Tau interaction via Ser356, advancing the understanding of IDP interactions and prompting re-evaluation 14-3-3/Tau mechanisms.

Language: Английский

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Tunable Stimuli-Responsive Module Based on α-Hydroxymethyl-α,β-Unsaturated Carbonyl Scaffold

Journal of Materials Chemistry B, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

The α-hydroxymethyl-α,β-unsaturated carbonyl (HMUC) scaffold represents a valuable framework for constructing nucleophile-responsive materials.

Language: Английский

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Design and application of a fluorescent probe for imaging of endogenous Bruton’s tyrosine kinase with preserved enzymatic activity

RSC Chemical Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Development of a novel fluorescent probe for endogenous BTK imaging using evobrutinib as scaffold. Evo-2 enabled real-time visualisation dynamics in living cells while preserving its enzymatic activity.

Language: Английский

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A modular click ligand-directed approach to label endogenous dopamine D1 receptors in live cells

Communications Chemistry, Journal Year: 2025, Volume and Issue: 8(1)

Published: April 11, 2025

Language: Английский

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