Fig. 3: OptoNBs can be deployed against multiple targets and used to control intracellular signaling.
From: Optogenetic control of protein binding using light-switchable nanobodies
a Superimposed nanobody structures with diverse binding modes: the anti-GFP minimizer nanobody in gray (PDB ID: 3G9A), anti-EGFR 7D12 in blue (PDB ID: 4KRL), anti-VGLUT Nb9 in beige (PDB ID: 5OCL), anti-Gelsolin Nb11 in pink (PDB ID: 4S10), and anti-CD38 MU551 in green (PDB ID: 5F1O). The CDRs, GG15 position, and AK74 position are highlighted in colors as indicated. b Light-induced translocation from nucleus to cytosol of LaM8, LaM4, and LaG9 OptoNBs with LOV insertion at GG15 or AK74. The change in cytosolic intensity from the original, dark-equilibrated value is shown. Error bars indicate mean ± SEM for n = 8, 4, 5, 8, 6, and 6 cells, respectively. c, d NIH3T3 cell lines harboring OptoNB-controlled Ras/Erk pathway activity using LaM8-GG15 (in c) and LaM8-AK74 (in d). Upper diagrams show lentiviral constructs expressed in each cell: membrane-localized mCherry-CAAX, OptoNB-SOScat, and a live-cell biosensor of Erk activity (ErkKTR-iRFP). Lower diagrams indicate mCherry and ErkKTR expression and localization for representative cells. Curves show the cytosolic-to-nuclear ratio of ErkKTR for a representative cell when pulsed with blue light (blue bars). Images are representative of three replicate experiments. Scale bars: 10 μm. Source data are available as a Source data file.
