Tyrosinase-Based Proximity Labeling in Living Cells and In Vivo

Journal of the American Chemical Society, Год журнала: 2024, Номер 146(11), С. 7515 - 7523

Опубликована: Март 6, 2024

Characterizing the protein constituents of a specific organelle and neighbors interest (POI) is essential for understanding function state networks associated with POI. Proximity labeling (PL) has emerged as promising technology efficient spatial proteomics. Nevertheless, most enzymes adopted PL still have limitations: APEX requires cytotoxic H2O2 activation thus poor in biocompatibility vivo application, BioID shows insufficient kinetics, TurboID suffers from high background biotinylation. Here, we introduce bacterial tyrosinase (BmTyr) new enzyme suitable H2O2-free, fast (≤10 min living cells), low-background tagging. BmTyr genetically encodable enables subcellular-resolved proteomics cells. We further designed strategy ligand-tethered PL, which unveiled surrounding proteome neurotransmitter receptor (Grm1 Drd2) its resident synapse live mouse brain. Overall, one that can improve expand PL-based applications discoveries.

Язык: Английский

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Recent advances in self‐immobilizing fluorescent probes for in vivo imaging

Smart Molecules, Год журнала: 2024, Номер 2(3)

Опубликована: Авг. 23, 2024

Abstract In situ precise detection of bioactive molecules with high sensitivity and spatiotemporal resolution is essential for studying physiological events disease diagnosis. The utilization versatile fluorescent probes in fluorescence imaging offers a powerful tool vivo biomarkers closely associated pathological conditions. However, the dynamic behavior leading to rapid clearance small molecule from regions interest severely compromises their potential imaging. Notably, self‐immobilizing that selectively recognize diseased tissues while improving retention enrichment enable accurate high‐fidelity this review, we aim summarize strategies employed recent advances performance precision using techniques. Lastly, discuss prospects challenges promote further development application more delicate probes.

Язык: Английский

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Selective fluorescent labeling of cellular proteins and its biological applications

Chemical Society Reviews, Год журнала: 2024, Номер 53(19), С. 9446 - 9489

Опубликована: Янв. 1, 2024

Proteins, which are ubiquitous in cells and critical to almost all cellular functions, indispensable for life. Fluorescence imaging of proteins is key understanding their functions within native milieu, as it provides insights into protein localization, dynamics, trafficking living systems. Consequently, the selective labeling target with fluorophores has emerged a highly active research area, encompassing bioorganic chemistry, chemical biology, cell biology. Various methods selectively tissues have been established continually being developed visualize characterize proteins. This review highlights findings reported since 2018, focus on small organic biological applications studying protein-associated events. We also discuss strengths weaknesses each approach utility

Язык: Английский

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Targeted Covalent Modification Strategies for Drugging the Undruggable Targets

Chemical Reviews, Год журнала: 2025, Номер unknown

Опубликована: Янв. 7, 2025

The term "undruggable" refers to proteins or other biological targets that have been historically challenging target with conventional drugs therapeutic strategies because of their structural, functional, dynamic properties. Drugging such undruggable is essential develop new therapies for diseases where current treatment options are limited nonexistent. Thus, investigating methods achieve drugging an important challenge in medicinal chemistry. Among the numerous methodologies drug discovery, covalent modification has emerged as a transformative strategy. attachment diverse functional molecules provides powerful platform creating highly potent and chemical tools well ability provide valuable information on structures dynamics targets. In this review, we summarize recent examples biomolecules development therapeutics overcome discovery challenges highlight how contribute toward particular, focus use chemistry drugs, identification, screening, artificial modulation post-translational modifications, cancer specific chemotherapies, nucleic acid-based therapeutics.

Язык: Английский

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Photoproximity labeling of endogenous receptors in the live mouse brain in minutes

Nature Chemical Biology, Год журнала: 2024, Номер unknown

Опубликована: Авг. 1, 2024

Язык: Английский

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N-Acyl-N-alkyl/aryl Sulfonamide Chemistry Assisted by Proximity for Modification and Covalent Inhibition of Endogenous Proteins in Living Systems

Accounts of Chemical Research, Год журнала: 2024, Номер unknown

Опубликована: Дек. 11, 2024

ConspectusSelective chemical modification of endogenous proteins in living systems with synthetic small molecular probes is a central challenge biology. Such has variety applications important for biological and pharmaceutical research, including protein visualization, functionalization, proteome-wide profiling enzyme activity, irreversible inhibition activity. Traditional chemistry selective cells largely relies on the high nucleophilicity cysteine residues to ensure target-selectivity site-specificity modification. More recently, lysine residues, which are more abundant surfaces, have attracted attention covalent proteins. However, it been difficult efficiently modify ε-amino groups side-chains, mostly (∼99.9%) protonated thus exhibit low at physiological pH. Our group revealed that

Язык: Английский

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Covalent functionalization of G protein-coupled receptors by small molecular probes

RSC Chemical Biology, Год журнала: 2025, Номер unknown

Опубликована: Янв. 1, 2025

In this review, we take a closer look at the most recent small molecular probes that covalently label G protein-coupled receptors.

Язык: Английский

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

Communications Chemistry, Год журнала: 2025, Номер 8(1)

Опубликована: Апрель 11, 2025

Язык: Английский

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Immobilizing the Endogenous 5-Hydroxytryptamine Transporter by Ligand-Directed and Proximity-Catalyzed Acyl Imidazole Chemistry for Affinity Chromatographic Analysis

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Май 3, 2025

The sensitivity, selectivity, and accuracy of immobilized protein-based methods are critically dependent on the strategies employed for protein immobilization. Compared with random immobilization approaches, site-specific covalent have emerged as promising alternatives, offering enhanced analytical performance. However, these typically require genetic modification target to incorporate a specific tag or prior purification protein, posing significant challenges immobilizing endogenous proteins. Herein, we address limitations by using 5-hydroxytryptamine transporter (5-HTT) model system. We designed probe conjugating fluvoxamine─a ligand 5-HTT─with either fluorescent reporter aminopropyl-modified silica gel through an alkyl linker containing acyl imidazole moiety reactive group. By introducing ligand-directed (LDAI) chemistry, achieved selective labeling 5-HTT in live cells without advance. Labeling experiments confirmed high specificity favorable kinetics LDAI ligands toward 5-HTT. Chromatographic analysis revealed improvements selectivity (5.7-50.9-fold), resolution (1.2-3.0-fold), sensitivity (1.0-12.0-fold) analyzing canonical when 5-HTT, compared His-tagged Halo-tagged recombinant This approach, substituting fluvoxamine other probe, can be generalized wide range proteins, versatile platform advanced techniques.

Язык: Английский

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In-brain synthesis of receptor-based protease sensors by coupling ligand-directed chemistry and click chemistry

Nature Synthesis, Год журнала: 2025, Номер unknown

Опубликована: Июнь 2, 2025

Язык: Английский

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