Light-Mediated Inhibition of Protein Synthesis

Abstract

The regulation of protein synthesis is vital for a host of cell biological processes, but investigating roles for protein synthesis have been hindered by the inability to selectively interfere with it. To inhibit protein synthesis with spatial and temporal control, we have developed a photo-releasable anisomycin compound, N-([6-bromo-7-hydroxycoumarin-4-yl]methyloxycarbonyl)anisomycin (Bhc-Aniso), that can be removed through exposure to UV light. The area of protein synthesis inhibition can be restricted to a small light-exposed region or, potentially, the volume of two-photon excitation if a pulsed IR laser is the light source. We have tested the compound's effectiveness with an in vitro protein-translation system, CHO cells, HEK293 cells, and neurons. The photo-released anisomycin can inhibit protein synthesis in a spatially restricted manner, which will enable the specific inhibition of protein synthesis in subsets of cells with temporal and spatial precision.

Technical Highlights of the Recent Literature

“…Global interference with translation is possible with cycloheximide, but spatial and temporal control of protein synthesis has proved elusive. In a recent report, Goard et al investigated whether photocaging the translational inhibitor anisomycin could be a practical means to achieve focused inhibition of protein synthesis. Photolabile groups have been extensively used to sequester effector molecules until UV-triggered uncaging; in this case anisomycin was caged with the addition of 4,5-xycoumarin-4-yl-methyl (Bhc), a photolabile group that demonstrated both stability and efficient uncaging in preliminary tests.…”

Von Ein‐ zu Zwei‐Photonen‐Sonden: photoaktivierbare Reagentien, Aktuatoren und Photoschalter

“…Bhc-Derivate wurden in verschiedenen Studien als zellmembrangängige Reagentien eingesetzt: für die intrazelluläre TP-Photolyse von cyclischen Nukleotid-Sekundärboten-stoffen; [49] für die Freisetzung von GABA in pyramidalen CA1-Zellen des Hippocampus; [50] für die Inhibierung der Proteinsynthese durch die In-vivo-Freisetzung von Anisomycin; [51] für die Photolysis von Bhc-abgeleiteten Progesteron-Regioisomeren; [52] und für die In-vivo-Aktivierung eines kompetitiven Inhibitors der induzierbaren Stickoxid-Synthase (iNOS). [53] Das wasserlçslichere 10 c (BCMACM) wurde für die Freisetzung eines cGMP-Analogons zur Auslçsung des Ca 2+ -Einstroms in CNG-transfizierte HEK293-Zellen verwendet.…”

A photochemical approach for controlled drug release in targeted drug delivery

“…The focus of this release method has mainly been on chemical and biological problems, such as the spatiotemporal control of cell signaling processes 34-36 , and it was only recently applied to drug delivery 8, 11-13 . The photocleavable linkers that are applicable for the current application are comprised of those based on o -nitrobenzyl (ONB) 33, 35, 37 , coumarin 35-36 , xanthene 38 , and benzophenone 39 . Figure 2 illustrates a schematic for controlled drug release triggered by a photochemical mechanism where a drug molecule attached to the photocleavable linker is released upon UV irradiation 8 .…”