Light-Activated Staudinger–Bertozzi Ligation within Living Animals

Shah, Lisa; Laughlin, Scott T.; Carrico, Isaac S.

doi:10.1021/jacs.5b13401

Cited by 69 publications

(57 citation statements)

References 34 publications

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“…Variants of the Staudinger reduction are some of the most bioorthogonal reactions reported to date, and have shown the ability to function in complex biological systems . We have demonstrated simple structural alterations to genetically encoded lysine protecting groups that rapidly accelerate deprotection by a Staudinger reduction/self‐immolation sequence, thus overcoming the most significant limitation—slow kinetics—while also improving deprotection yield.…”

Section: Resultsmentioning

confidence: 99%

“…This approach allowed us to control protein subcellular localization via multiple mechanisms (controlling NLS function and controlling SUMOylation) and to target proteins to different subcellular locations. The methodology demonstrated here is generally applicable to other proteins, cell types, and model organisms . In contrast to light as another fast trigger of protein function, our small‐molecule‐triggered system does not require specialized equipment, is not affected by tissue opaqueness, and does not interfere with fluorescent reporters .…”

Section: Resultsmentioning

confidence: 99%

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Phosphine‐Activated Lysine Analogues for Fast Chemical Control of Protein Subcellular Localization and Protein SUMOylation

et al. 2019

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The Staudinger reduction and its variants have exceptional compatibility with live cells but can be limited by slow kinetics. Herein we report new small‐molecule triggers that turn on proteins through a Staudinger reduction/self‐immolation cascade with substantially improved kinetics and yields. We achieved this through site‐specific incorporation of a new set of azidobenzyloxycarbonyl lysine derivatives in mammalian cells. This approach allowed us to activate proteins by adding a nontoxic, bioorthogonal phosphine trigger. We applied this methodology to control a post‐translational modification (SUMOylation) in live cells, using native modification machinery. This work significantly improves the rate, yield, and tunability of the Staudinger reduction‐based activation, paving the way for its application in other proteins and organisms.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Phosphine‐Activated Lysine Analogues for Fast Chemical Control of Protein Subcellular Localization and Protein SUMOylation

et al. 2019

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“…Reactions that perform well in vivo can therefore be considered the gold-standard of click reactions. [13][14] Especially the photochemically controlled strain-promoted azide alkyne coupling (photo-SPAAC) and the 1,3-dipolar cycloaddition between a nitrile imine generated from a tetrazole under light irradiation and an alkene (Nitrile Imine mediated Tetrazole Ene Cycloaddition, NITEC) are both often considered to be bioorthogonal. 2,[15][16] However, despite their asserted bioorthogonality, specific limitations apply when these reactions are employed.…”

Section: 4mentioning

confidence: 99%

Visible Light-Induced Ligation via o-Quinodimethane Thioethers

et al. 2018

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We introduce a photocaged diene system ( o-quinodimethane thioethers) based on o-methylbenzaldehydes ( o-MBAs) that can be activated with visible light. The pioneered system is accessible in a single step from commercially available starting materials in excellent yields. Variable synthetic handles can be attached to the photocaged diene, often without elaborate protecting group chemistry. Full conversion of various o-methylbenzaldehydes to the Diels-Alder adduct is achieved in the presence of maleimides under catalyst-free conditions triggered by visible light irradiation with LEDs under flow conditions. Unlike the previously reported UV-induced ligation of o-quinodimethanes, the reaction can be conducted both in organic solvents and in aqueous solution. We further demonstrate the ability of the photocaged dienes to ligate two polymer blocks by visible light. The [4+2] nature of the reaction makes it a powerful orthogonal ligation platform.

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“…2000) (Fig. 10 a ), as well as reactions based on azobenzene or caged phosphine reagents that can be activated with light (Shah et al . 2016; Szymanski et al .…”

Section: Molecular Engineering Toolbox For Complex Biological Samplesmentioning

confidence: 99%

A molecular engineering toolbox for the structural biologist

Debelouchina

Muir

2017

Quart. Rev. Biophys.

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Exciting new technological developments have pushed the boundaries of structural biology, and have enabled studies of biological macromolecules and assemblies that would have been unthinkable not long ago. Yet, the enhanced capabilities of structural biologists to pry into the complex molecular world have also placed new demands on the abilities of protein engineers to reproduce this complexity into the test tube. With this challenge in mind, we review the contents of the modern molecular engineering toolbox that allow the manipulation of proteins in a site-specific and chemically well-defined fashion. Thus, we cover concepts related to the modification of cysteines and other natural amino acids, native chemical ligation, intein and sortase-based approaches, amber suppression, as well as chemical and enzymatic bio-conjugation strategies. We also describe how these tools can be used to aid methodology development in X-ray crystallography, nuclear magnetic resonance, cryo-electron microscopy and in the studies of dynamic interactions. It is our hope that this monograph will inspire structural biologists and protein engineers alike to apply these tools to novel systems, and to enhance and broaden their scope to meet the outstanding challenges in understanding the molecular basis of cellular processes and disease.

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Light-Activated Staudinger–Bertozzi Ligation within Living Animals

Cited by 69 publications

References 34 publications

Phosphine‐Activated Lysine Analogues for Fast Chemical Control of Protein Subcellular Localization and Protein SUMOylation

Phosphine‐Activated Lysine Analogues for Fast Chemical Control of Protein Subcellular Localization and Protein SUMOylation

Visible Light-Induced Ligation via o-Quinodimethane Thioethers

A molecular engineering toolbox for the structural biologist

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