Fig. 5: In situ TXTL synthesis of Min proteins induces coherent long-range mass oscillations across the SLB-Sensor system. | Communications Biology

Fig. 5: In situ TXTL synthesis of Min proteins induces coherent long-range mass oscillations across the SLB-Sensor system.

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

Fig. 5

a Cell-free synthesis of MinD (linear P70a-minD) into the QCMD modules on top of SLBs. The cartoon shows a very simplified model of the synthesized MinD binding kinetics when the reaction is in contact with either a pure DOPC SLB, a DOPC/DOPG SLB, or a DOPC/CL SLB. b Adsorption kinetics of MinD with either a pure DOPC SLB, a DOPC/DOPG SLB, or a DOPC/CL SLB. The curves are labeled with a few symbols, the lines are experimental points. c Number of oscillation peaks within the first 2 hours as a function of the relative DOPG concentration during SALB for a DOPE and DOPC Base SLB. d Duration of oscillations as a function of the relative DOPG concentration during SALB for a DOPE and DOPC Base SLB. e Number of oscillation peaks within the first 2 hours as a function of the relative CL concentration during SALB for a DOPE and DOPC Base SLB. f Duration of oscillations as a function of the relative CL concentration during SALB for a DOPE and DOPC Base SLB. g Cell-free synthesis of MinD with either MinE (linear P70a-minE) or MinC (linear P70a-minC) into the QCMD modules on top of SLBs. h Adsorption kinetics of MinD (P70a-minD, 10 nM), MinDE (P70a-minD, 10 nM and P70a-minE, 0.1 nM), and MinCD (P70a-minD, 10 nM and P70a-minC, 5 nM), on a 10% mol ratio DOPG/DOPE SLB. The left inlet shows a close-up of the oscillations during the first 2.4 hours. The right inlet shows a close-up of the oscillations between t = 5 hours and t = 11 hours. The curves are labeled with a few symbols, the lines are experimental points. i Duration of oscillations as a function of P70a-minE concentration in the TXTL reaction for a DOPG/DOPE SLB, a DOPG/DOPC SLB, and an E. coli lipids (ECL) SLBs. j Adsorption kinetics of a MinDE (P70a-minD, 10 nM and P70a-minE, 0.1 nM) on a 10% mol ratio DOPG/DOPE SLB with a range of PEG5k-PE added during SALB. The curves are labeled with a few symbols, the lines are experimental points. Each measurement is represented as an individual data point.

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