Author/Authors :
Michael Goard، نويسنده , , Girish Aakalu، نويسنده , , Olesya D. Fedoryak، نويسنده , , Carlo Quinonez، نويسنده , , Jamii St. Julien، نويسنده , , Stephen J. Poteet، نويسنده , , Erin M. Schuman، نويسنده , , Timothy M. Dore، نويسنده ,
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.