Fig. 3: In situ cell-free synthesized MscL and AH-eGFP exhibit their natural phospholipid affinities in ‘pure’ and ‘mixed’ SLBs. | Communications Biology

Fig. 3: In situ cell-free synthesized MscL and AH-eGFP exhibit their natural phospholipid affinities in ‘pure’ and ‘mixed’ SLBs.

From: Cell-free expression with a quartz crystal microbalance enables rapid, dynamic, and label-free characterization of membrane-interacting proteins

Fig. 3

a MscL or AH-eGFP is synthesized into the QCMD module on top of either a PC-rich EggPC SLB or a PE-rich E. coli lipids (ECL) SLB. The rightmost cartoon demonstrates the expected protein-lipid preferences of both proteins. b, c Adsorption kinetics of blank (P70a-T7rnap, 0.15 nM), AH-eGFP (P70a-T7rnap, 0.15 nM, T7p14-ah-egfp, 5 nM), and MscL (P70a-T7rnap, 0.15 nM, T7p14-mscL, 5 nM) synthesizing TXTL reactions in contact with an ECL SLB and an EggPC SLB respectively. d The frequency changes after 8 hours of incubating either blank (P70a-T7rnap, 0.15 nM), AH-eGFP (P70a-T7rnap, 0.15 nM, T7p14-ah-egfp, 5 nM), or MscL (P70a-T7rnap, 0.15 nM, T7p14-mscL, 5 nM) TXTL reactions in ECL–EggPC Mixed SLBs. The x-axis percentage indicates the concentration of EggPC (and ECL) relative to the working concentration from Table S1. For example, 40% EggPC means that the Lipid-IPA mix contains 0.52 mM of EggPC phospholipids (40% of 1.3 mM) and 1.8 mM of ECL phospholipids (60% of 3 mM). All of the performed replicates are shown in the figure as individual data points.

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