Controlling protein reaction in cells with visible light- our preprint

We have just posted on bioRxiv a preprint on our rapid-reacting protein module that is now activatable in cells with light: “Visible light-induced specific protein reaction delineates early stages of cell adhesion”. I am keen to get your feedback as we develop this approach, either in the comments below here, at the bioRxiv site, or e-mail me directly. We have caged the reactive lysine of SpyCatcher003 with a hydroxycoumarin-containing unnatural amino acid. So in the dark there is no reaction with SpyTag003. Upon illumination with laser or wide-field at 405 nm (in the visible range), covalent reaction with SpyTag003 starts rapidly. We applied this control of reactivity to investigate the earliest steps as cells start to build their adhesions, linking the extracellular matrix with the cytoskeleton and sensing forces. We can see effects from talin reconstitution in less than 30 seconds and follow by microscopy the order of recruitment of adhesion components. This is being led by Rolle Rahikainen with Paula Turkki and Vesa Hyt?nen, based on work started by Susan Vester. The unnatural amino acid expertise comes from Chasity P. Janosko and Alexander Deiters. Hopefully this should be a helpful addition to the optogenetic toolbox.

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