The value of coimmunoprecipitation (Co-IP) assays in drug discovery DOI

Zhongtian Cai,

Danni Wang, Zekun Li

et al.

Expert Opinion on Drug Discovery, Journal Year: 2025, Volume and Issue: unknown, P. 1 - 14

Published: April 28, 2025

Co-IP assays are well-established technologies widely applicated for investigating the mechanisms underlying protein-protein interactions and identifying interaction modulators. These play an important role in elucidating complex networks of protein critical cellular functions. This review covers a technical protocol standard Co-IP. The research contents conclusions modulators summarized. Finally, three derivations introduced. Literature was surveyed from original publications, sources, PubMed clinical trials through 14 April 2025. To perform successfully, researchers must consider selection specific antibody, remission nonspecific binding detection limitations transient or weak interactions. offer several advantages over tandem affinity purification pull-down methods, particularly their applicability to primary cells. allows study PPIs natural environment. Conventional often struggle detect can suffer contamination. However, advancements techniques address these challenges, enhancing sensitivity specificity, enabling subtle while distinguishing events. makes powerful tool exploring dynamics living systems.

Language: Английский

Photocatalytic Late-Stage C–H Functionalization DOI
Peter Bellotti, Huan‐Ming Huang,

Teresa Faber

et al.

Chemical Reviews, Journal Year: 2023, Volume and Issue: 123(8), P. 4237 - 4352

Published: Jan. 24, 2023

The emergence of modern photocatalysis, characterized by mildness and selectivity, has significantly spurred innovative late-stage C–H functionalization approaches that make use low energy photons as a controllable source. Compared to traditional strategies, photocatalysis paves the way toward complementary and/or previously unattainable regio- chemoselectivities. Merging compelling benefits with workflow offers potentially unmatched arsenal tackle drug development campaigns beyond. This Review highlights photocatalytic strategies small-molecule drugs, agrochemicals, natural products, classified according targeted bond newly formed one. Emphasis is devoted identifying, describing, comparing main mechanistic scenarios. draws critical comparison between established ionic chemistry photocatalyzed radical-based manifolds. aims establish current state-of-the-art illustrate key unsolved challenges be addressed in future. authors aim introduce general readership functionalization, specialist practitioners evaluation methodologies, potential for improvement, future uncharted directions.

Language: Английский

Citations

324
Advances in theE → ZIsomerization of Alkenes Using Small Molecule Photocatalysts DOI
Tomáš Neveselý,

Max Wienhold,

John J. Molloy

et al.

Chemical Reviews, Journal Year: 2021, Volume and Issue: 122(2), P. 2650 - 2694

Published: Aug. 27, 2021

Geometrical E → Z alkene isomerization is intimately entwined in the historical fabric of organic photochemistry and enjoying a renaissance (Roth et al. Angew. Chem., Int. Ed. Engl. 1989 28, 1193–1207). This consequence fundamental stereochemical importance Z-alkenes, juxtaposed with frustrations thermal reactivity that are rooted microscopic reversibility. Accessing excited state paradigms allow this latter obstacle to be circumnavigated by exploiting subtle differences photophysical behavior substrate product chromophores: provides molecular basis for directionality. While direct irradiation operationally simple, photosensitization via selective energy transfer enables augmentation repertoire include substrates not directly photons. Through sustained innovation, an impressive portfolio tailored small molecule catalysts range triplet energies now widely available facilitate contra-thermodynamic thermo-neutral reactions generate Z-alkene fragments. review intended serve as practical guide covering geometric alkenes enabled catalysis from 2000 2020, logical sequel excellent treatment Dugave Demange (Chem. Rev. 2003 103, 2475–2532). The mechanistic foundations underpinning selectivity discussed together induction models rationales explain counterintuitive directionality these processes which very distinguish product. Implications subsequent stereospecific transformations, application total synthesis, regioselective polyene isomerization, spatiotemporal control pre-existing configuration broader sense discussed.

Language: Английский

Citations

309
μMap-Red: Proximity Labeling by Red Light Photocatalysis DOI

Benito F. Buksh,

Steve D. Knutson, James V. Oakley

et al.

Journal of the American Chemical Society, Journal Year: 2022, Volume and Issue: 144(14), P. 6154 - 6162

Published: April 1, 2022

Modern proximity labeling techniques have enabled significant advances in understanding biomolecular interactions. However, current tools primarily utilize activation modes that are incompatible with complex biological environments, limiting our ability to interrogate cell- and tissue-level microenvironments animal models. Here, we report μMap-Red, a platform uses red-light-excited Sn

Language: Английский

Citations

73
Tracking chromatin state changes using nanoscale photo-proximity labelling DOI
Ciaran P. Seath, Antony J. Burton,

Xuemeng Sun

et al.

Nature, Journal Year: 2023, Volume and Issue: 616(7957), P. 574 - 580

Published: April 5, 2023

Language: Английский

Citations

58
Multiscale photocatalytic proximity labeling reveals cell surface neighbors on and between cells DOI
Zhi Lin, Kaitlin Schaefer, Irene Lui

et al.

Science, Journal Year: 2024, Volume and Issue: 385(6706)

Published: July 18, 2024

Proximity labeling proteomics (PLP) strategies are powerful approaches to yield snapshots of protein neighborhoods. Here, we describe a multiscale PLP method with adjustable resolution that uses commercially available photocatalyst, Eosin Y, which upon visible light illumination activates different photo-probes range radii. We applied this platform profile neighborhoods the oncogenic epidermal growth factor receptor and orthogonally validated more than 20 neighbors using immunoassays AlphaFold-Multimer prediction. further profiled cell-cell synapses induced by bispecific T cell engagers chimeric antigen cells. This integrated maps local distal networks on between surfaces, will aid in systematic construction surface interactome, revealing horizontal signaling partners reveal new immunotherapeutic opportunities.

Language: Английский

Citations

24
Photocatalytic Activation of Aryl(trifluoromethyl) Diazos to Carbenes for High-Resolution Protein Labeling with Red Light DOI
David C. Cabanero, Stavros K. Kariofillis, Andrew C. Johns

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(2), P. 1337 - 1345

Published: Jan. 2, 2024

State-of-the-art methods in photoproximity labeling center on the targeted generation and capture of short-lived reactive intermediates to provide a snapshot local protein environments. Diazirines are current gold standard for high-resolution proximity labeling, generating aryl(trifluoromethyl) carbenes. Here, we present method access carbenes from stable diazo source via tissue-penetrable, deep red near-infrared light (600–800 nm). The operative mechanism this activation involves Dexter energy transfer photoexcited osmium(II) photocatalysts diazo, thus revealing an carbene. preferences probe with amino acids studied, showing high reactivity toward heteroatom−H bonds. Upon synthesis biotinylated probe, studies conducted native proteins as well conjugated Os photocatalyst. Finally, demonstrate that conjugation inhibitor photocatalyst also enables selective presence spectator achieves specific membrane surface mammalian cells two-antibody photocatalytic system.

Language: Английский

Citations

21
Current advances in photocatalytic proximity labeling DOI
Steve D. Knutson,

Benito F. Buksh,

Sean W. Huth

et al.

Cell chemical biology, Journal Year: 2024, Volume and Issue: 31(6), P. 1145 - 1161

Published: April 24, 2024

Language: Английский

Citations

16
Spatiotemporal-resolved protein networks profiling with photoactivation dependent proximity labeling DOI Creative Commons

Yansheng Zhai,

Xiaoyan Huang, Keren Zhang

et al.

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: Aug. 20, 2022

Abstract Enzymatic-based proximity labeling approaches based on activated esters or phenoxy radicals have been widely used for mapping subcellular proteome and protein interactors in living cells. However, are poorly reactive which leads to a wide radius generated by peroxide treatment may disturb redox-sensitive pathways. Herein, we report photoactivation-dependent (PDPL) method designed genetically attaching photosensitizer miniSOG of interest. Triggered blue light tunned irradiation time, singlet oxygen is generated, thereafter enabling spatiotemporally-resolved aniline probe histidine residues. We demonstrate its high-fidelity through organelle-specific proteomes. Side-by-side comparison PDPL with TurboID reveals more specific deeper proteomic coverage PDPL. further apply the disease-related transcriptional coactivator BRD4 E3 ligase Parkin, discover previously unknown interactors. Through over-expression screening, two unreported substrates Ssu72 SNW1 identified whose degradation processes mediated ubiquitination-proteosome pathway.

Language: Английский

Citations

45
Bioorthogonal masked acylating agents for proximity-dependent RNA labelling DOI
Shubhashree Pani, Tian Qiu, Kaitlin Kentala

et al.

Nature Chemistry, Journal Year: 2024, Volume and Issue: 16(5), P. 717 - 726

Published: April 9, 2024

Language: Английский

Citations

15
Intracellular Synthesis of Indoles Enabled by Visible-Light Photocatalysis DOI Creative Commons
Cinzia D’Avino, Sara Gutiérrez, Max J. Feldhaus

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(5), P. 2895 - 2900

Published: Jan. 26, 2024

Performing abiotic synthetic transformations in live cell environments represents a new, promising approach to interrogate and manipulate biology uncover new types of biomedical tools. We now found that photocatalytic bond-forming reactions can be added the toolbox bioorthogonal chemistry. Specifically, we demonstrate exogenous styryl aryl azides converted into indoles inside living mammalian cells under conditions.

Language: Английский

Citations

14