Fig. 5: Bioluminescence-activated membrane recruitment in synthetic cells.
From: Synthetic cells with self-activating optogenetic proteins communicate with natural cells
a A block diagram of the Gaussia luciferase (Gluc)-iLID fusion protein elements. Below, a schematic representation of protein recruitment to the synthetic cell membrane by hetero-dimerization of the fusion protein Gluc-iLID with RFP-sspB. b Microscopy image of luciferase light emission from synthetic cells with membrane-bound Gluc-EL222 after coelenterazine addition. This experiment was reproduced n = 5 times. c (i) Membrane recruitment of RFP-sspB with iLID or Gluc-iLID using 488 nm laser illumination or by addition of coelenterazine to activate the bioluminescent reaction. RFP intensity is normalized to the average intensity of each cell in the dark conditions. Data is expressed as a mean ± s.e.m. Welch’s two-tailed t-test P-values; **p = 0.0092, ***p = 0.0001, for comparison of Gluc-iLID vs. iLID activated by laser and iLID supplemented with coelenterazine at t = 6 min, respectively. (ii) Representative single-cell images of RFP-sspB recruitment to a synthetic cell with membrane-bound Gluc-iLID in the dark and after two and four doses of 0.2 nmol coelenterazine (Top row). Bottom row displays the fluorescence of the synthetic cell’s DOPE-Cy5 lipid that composed 0.5 mol% of the membrane (n = 3 for iLID + coelenterazine, n = 11 for iLID + laser, n = 14 for Gluc-iLID + coelenterazine). Source data are provided as a Source Data file.
