Computational design and experimental characterization of a photo-controlled mRNA-cap guanine-N7 methyltransferase

Abstract

The spatial and temporal control of gene expression at the post-transcriptional level is essential in eukaryotic cells and developing multicellular organisms. In recent years optochemical and optogenetic tools have enabled...

Visible Light Control over the Cytolytic Activity of a Toxic Pore-Forming Protein

“…However, there are only a few examples in which the photoswitch is covalently attached to a protein. As such, tetra- ortho -fluorinated azobenzene-based amino acids were encoded into mammalian cells via genetic code expansion enabling incorporation of the switch in vivo . , Furthermore, both fluorinated and chlorinated azobenzenes were tethered to ionotropic receptors to modulate their activity. , Visible light-responsive photoswitches with two attachment points, so-called cross-linkers, have been introduced as a staple to control the secondary structure of bioactive short peptides. ,,− However, to the best of our knowledge, there is only one example in which the visible-light responsive switch (based on the tetra- ortho -methoxy azobenzene core) was utilized to control a protein function through cross-linking the binding site of a guanine-N7 methyltransferase to achieve visible-light mediated methylation of the 5′ cap of mRNA . Nevertheless, none of the described examples exhibited pharmacological (cytolytic or cytotoxic) activity, specifically, when targeting cancer cells.…”

“… 58 , 75 , 83 − 85 However, to the best of our knowledge, there is only one example in which the visible-light responsive switch (based on the tetra- ortho -methoxy azobenzene core) was utilized to control a protein function through cross-linking the binding site of a guanine-N7 methyltransferase to achieve visible-light mediated methylation of the 5′ cap of mRNA. 86 Nevertheless, none of the described examples exhibited pharmacological (cytolytic or cytotoxic) activity, specifically, when targeting cancer cells. Therefore, in this study, we report for the first time the fully visible-light-mediated control of protein toxicity with a water-soluble molecular photoswitch.…”