Multifunctional Photo-Cross-Linking Probes: From Target Protein Searching to Imaging Applications

Kozoriz, Kostiantyn; Shkel, Olha; Hong, Kyung Tae; Kim, Dong Hoon; Kim, Yun Kyung; Lee, Jung Hoon

doi:10.1021/acs.accounts.2c00505

Cited by 14 publications

(9 citation statements)

References 66 publications

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“…The most conventional strategy for introducing BODIPY tags into protein structures involves the direct conjugation of reactive fluorophores (e.g., N -hydroxysuccinimides, maleimides) to nucleophilic amino acids (e.g., lysines and cysteines, respectively). , Despite being an effective approach to label proteins with BODIPY groups, these reactions are not site specific and yield heterogeneous mixtures with a diverse range of bioactivity profiles. The groups of Lee and Holland have recently reported photoactivatable fluorophores (e.g., aryl ketones and aryl azides, respectively) for light-controlled insertion of BODIPY moieties into proteins. , These reactions are fast and proceed with reasonable yields, but their labeling efficiency is also highly dependent on the accessibility of reactive residues.…”

Section: Resultsmentioning

confidence: 99%

“…The groups of Lee and Holland have recently reported photoactivatable fluorophores (e.g., aryl ketones and aryl azides, respectively) for light-controlled insertion of BODIPY moieties into proteins. 44,45 These reactions are fast and proceed with reasonable yields, but their labeling efficiency is also highly dependent on the accessibility of reactive residues. Alternatively, genetic code reprogramming using the amber suppression technology provides a means to reassign the amber stop codon to an unnatural amino acid and introduce an exogenous chemical moiety (e.g., fluorophore) at a single, specific position of the protein structure.…”

Section: Chemical Synthesis and Spectral Characterization Ofmentioning

confidence: 99%

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Inserting “OFF-to-ON” BODIPY Tags into Cytokines: A Fluorogenic Interleukin IL-33 for Real-Time Imaging of Immune Cells

Reese,

de Moliner,

Mendive-Tapia

et al. 2023

ACS Cent. Sci.

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The essential functions that cytokine/immune cell interactions play in tissue homeostasis and during disease have prompted the molecular design of targeted fluorophores to monitor their activity in real time. Whereas activatable probes for imaging immune-related enzymes are common, many immunological functions are mediated by binding events between cytokines and their cognate receptors that are hard to monitor by live-cell imaging. A prime example is interleukin-33 (IL-33), a key cytokine in innate and adaptive immunity, whose interaction with the ST2 cell-surface receptor results in downstream signaling and activation of NF-κB and AP-1 pathways. In the present work, we have designed a chemical platform to site-specifically introduce OFF-to-ON BODIPY fluorophores into full cytokine proteins and generate the first nativelike fluorescent analogues of IL-33. Among different incorporation strategies, chemical aminoacylation followed by bioorthogonal derivatization led to the best labeling results. Importantly, the BODIPY-labeled IL-33 derivatives�unlike IL-33-GFP constructs�exhibited ST2-specific binding and downstream bioactivity profiles comparable to those of the wild-type interleukin. Real-time fluorescence microscopy assays under no wash conditions confirmed the internalization of IL-33 through ST2 receptors and its intracellular trafficking through the endosomal pathway. We envision that the modularity and versatility of our BODIPY labeling platform will facilitate the synthesis of minimally tagged fluorogenic cytokines as the next generation of imaging reagents for real-time visualization of signaling events in live immune cells.

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Section: Resultsmentioning

confidence: 99%

Section: Chemical Synthesis and Spectral Characterization Ofmentioning

confidence: 99%

Inserting “OFF-to-ON” BODIPY Tags into Cytokines: A Fluorogenic Interleukin IL-33 for Real-Time Imaging of Immune Cells

Reese,

de Moliner,

Mendive-Tapia

et al. 2023

ACS Cent. Sci.

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“…SAR or structural information on the protein−ligand complex can identify potential attachment sites. The most common photoreactive groups are aryl azides, benzylic and aliphatic azirines, and benzophenones, which react to the corresponding nitrene (44), carbene (49), or ketyl radical (52), respectively (Figure 13). 47 In addition to the generation of reactive intermediates, photochemical electrophile generation approaches have also been reported.…”

Section: Photoaffinity and Photocatalytic Proximity Labelingmentioning

confidence: 99%

“…47 In addition to the generation of reactive intermediates, photochemical electrophile generation approaches have also been reported. 49 Photoreactive groups should be bioorthogonal and stable in the dark. Under consideration of their molecular size, reactivity, synthetic accessibility, and activation wavelength, each photoreactive group displays its own advantages and disadvantages.…”

Section: Photoaffinity and Photocatalytic Proximity Labelingmentioning

confidence: 99%

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Photochemistry in Medicinal Chemistry and Chemical Biology

Zhu,

Empel,

Pelliccia

et al. 2024

J. Med. Chem.

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Photochemistry has emerged as a transformative force in organic chemistry, significantly expanding the chemical space accessible for medicinal chemistry. Light-induced reactions enable the efficient synthesis of intricate organic structures and have found applications throughout the different stages of the drug discovery and development processes. Moreover, photochemical techniques provide innovative solutions in chemical biology, allowing precise spatiotemporal drug activation and targeted delivery. In this Perspective, we highlight the already numerous remarkable applications and the even more promising future of photochemistry in medicinal chemistry and chemical biology. ■ SIGNIFICANCE• Medicinal chemistry and chemical biology can highly benefit from the recent developments in the field of photochemistry in organic synthesis. • The most relevant applications of photochemistry and photocatalysis in the different stages of the drug discovery process have been highlighted. • With this Perspective, we would like to point out the potential and challenges of photochemistry in chemical biology applications.

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Fluorescent Labeling of a Target Protein with an Alkyl Diazirine Photocrosslinker Bearing a Cinnamate Moiety

Nakashima,

Iwanabe,

Tanimoto

et al. 2024

Chemistry An Asian Journal

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A novel fluorogenic alkyl diazirine photocrosslinker bearing an o‐hydroxycinnamate moiety has been developed for identification of the targets of bioactive molecules. The o‐hydroxycinnamate moiety can be converted to the corresponding 7‐hydroxycoumarin derivative, which should be created on the interacting site within the photocaptured target protein. The label yield and fluorescence intensity have been immensely improved in comparison with our previous aromatic crosslinkers to facilitate target identification in small quantities.

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Multifunctional Photo-Cross-Linking Probes: From Target Protein Searching to Imaging Applications

Abstract: Acc. Chem. Res. XXXX, XXX, XXX−XXX "minimalist tag" containing a series of bio-orthogonal tags. A set of aff inity-based probes for BRD4 inhibitors was created and investigated for interactome studies, resulting in novel off-target discovery.

Cited by 14 publications

References 66 publications

Inserting “OFF-to-ON” BODIPY Tags into Cytokines: A Fluorogenic Interleukin IL-33 for Real-Time Imaging of Immune Cells

Inserting “OFF-to-ON” BODIPY Tags into Cytokines: A Fluorogenic Interleukin IL-33 for Real-Time Imaging of Immune Cells

Photochemistry in Medicinal Chemistry and Chemical Biology

Fluorescent Labeling of a Target Protein with an Alkyl Diazirine Photocrosslinker Bearing a Cinnamate Moiety

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