Fig. 1: LCST-type phase behavior of the E. coli LplA enzyme protein in vitro.

a LplA at a concentration of 500 μM undergoes LCST-type sol-gel phase transition in vitro. b Scanning electron microscopy (SEM) image of LplA gel network. The image is representative of three independent experiments. c Normalized FRAP analysis of LplA-mEGFP condensates in a Tris buffer (pH 7.4) with 100 mM NaCl and 10% Dextran 70. Images are representative of two independent experiments. FRAP data are expressed as the mean ±s.d. for n = 5 condensates. d The thermo-responsive sol-gel reversible cycle of LplA characterized using turbidity at 600 nm. Data are representative of three independent experiments. e Rheological characterization of LplA hydrogel in temperature sweep mode. LplA sample is prepared in a Tris buffer (pH 7.4) with a protein concentration at 1 mM. Temperature sweeps were performed at 1.25 rad/s frequency and 0.1% strain. Data are representative of three independent experiments. f Phase diagrams of LplA at varying salt and protein concentrations in the absence of crowding agents. 50 mM Tris (pH 7.4) was used for solution buffering. Phase separation is determined using turbidity at 600 nm (at 30 °C). Data are representative of two independent experiments. g A summary of the phase behavior at room temperature of 1 mM LplA under different solution conditions. Data are representative of two independent experiments. h Small-angle X-ray scattering (SAXS) profiles of LplA at a concentration of 500 μM in a Tris buffer at different temperatures. Curves were shifted for clarity. The details of the correlation peaks (indicated by the dashed box in the left image) are displayed in the right image. The characteristic distances of the labeled peaks are ① 8.92 nm, ② 7.20 nm, ③ 6.16 nm and ④ 4.55 nm. Data are representative of two independent experiments. Source data are provided as a Source Data file.