Fig. 3
From: Artificial photosynthetic cell producing energy for protein synthesis
Self-constituting protein synthesis in artificial photosynthetic cells. a Schematics of self-constituting protein synthesis. The numbers indicate the order of reactions; ➀: adenosine triphosphate (ATP) synthesis, ➁: aminoacylation of transfer RNA (tRNA) by aminoacyl-tRNA synthetase (ARS), ➂: translation by ribosomes (Rbs), ➃: de novo bacteriorhodopsin (bR) synthesis, and ➄: de novo Fo synthesis. b Light-induced bR-GFP synthesis in giant unilamellar vesicles (GUVs). Bar: 10 µm. c Membrane localization of bR. The bRs synthesized in the PURE system with or without liposomes were fractionated by ultra-centrifugation with sucrose cushion. The percentages of bR in each fraction (%Frac.) are indicated at the bottom of the gels. d Proton-pump activity of bR synthesized in the PURE system. The measurement was performed as in Fig. 1c. The reaction times of protein synthesis are indicated by different colors. The white and gray areas represent light ON and OFF, respectively. e Enhanced artificial organelle by de novo bR. Wild-type (bRwt) or mutant (bRmut) bRs were photosynthesized in the PURE system containing bRFoF1-PLs. The ATP concentrations at the time 2 h was measured and converted into ATP per proteoliposome (PL). The value of the de novo bRwt-containing PL was normalized to that of the de novo bRmut-containing PL. ***P < 0.001. f Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the de novo photosynthesized bR. g Light-driven ATP synthesis by PLs consist of cell-free synthesized Fo. Wild-type (awt) or mutant (amut) a-subunit protein was synthesized together with b- and c-subunits in the PURE system containing purified F1 and bR-PLs. The measured ATP concentrations were converted into the produced ATP per PL. h Enhanced artificial organelle by de novo Fo. awt or amut was photosynthesized together with b- and c-subunits in the presence of purified F1 and bRFoF1-PLs. The ATP concentrations at the time 3 h was measured and converted into ATP per PL. The value of the de novo awt-containing PL was normalized by that of the de novo amut-containing PL. **P < 0.01. i SDS-PAGE analysis of the de novo photosynthesized Fo. P values were from two-side t-test. All experiments were performed at least three times and their means and S.D. are shown. Source data are provided as a Source Data file
