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

Abstract

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.

Small Molecule Control of Morpholino Antisense Oligonucleotide Function through Staudinger Reduction

“…In designing the linker, we utilized a carbamate leaving group, as it is stable to physiological conditions and well-established for self-immolation. , Furthermore, the benzylic position of the linker was substituted, as this has been shown to accelerate self-immolation by stabilizing the partial positive charge that develops during elimination . These design considerations are supported by recent comparisons of azidobenzyl carbamates as phosphine-removable protecting groups in biological settings. , Aside from the substituted p -azidobenzyl carbamate core, we functionalized the linker with N -hydroxysuccinimide (NHS) ester and chloroacetamide handles for cyclization of MOs modified at the termini with the proper amino and thiol handles.…”

“…The linker was synthesized in 10 steps from commercially available 4-aminoacetophenone ( 1 ) (Figure A). Diazotization and azide substitution of 1 yielded the aryl azide 2 . Bromination of 2 provided the α-bromo ketone 3 , which was then transformed into the corresponding primary amine 4 utilizing hexamine in a Delépine reaction.…”

“…Furthermore, they do not require irradiation equipment and they can be easily applied to large numbers of embryos in parallel. Various bioorthogonal small molecule pairs have been proven to be useful probes, enabling studies into biological processes without perturbing the endogenous system. − The Staudinger reduction and its variants are among the most bioorthogonal reactions and have consistently shown success in various demanding biological environments. − Based on their biorthogonality and history of use in vivo , we selected the azide/phosphine pair to generate a cMO linker that is cleaved through a Staudinger reduction-induced self-immolation.…”

Occupational airborne contact urticaria, anaphylaxis and asthma in farmers and agronomists due to Bruchus pisorum

“…According to Ministry of Agriculture regulations at the time of the study, peas had been treated only with aluminum phosphide EPA, ISO. 21 Magnesium phosphate was the pesticide used in previous years. The difference is that aluminum phosphate is a muricide and it was necessary to control voles that were transmitting tularemia in our region.…”

“…Our patients had previously handled seed pea for years and had used different insecticides, usually phosphates, for pest control. According to Ministry of Agriculture regulations at the time of the study, peas had been treated only with aluminum phosphide EPA, ISO 21 . Magnesium phosphate was the pesticide used in previous years.…”