Integral and interacting membrane proteins (IIMPs) are crucial biomolecules involved in various life processes, yet their characterization has been hindered by difficulties in purifying and labeling them. This study introduces a novel approach combining cell-free transcription-translation with quartz crystal microbalance with dissipation (TXTL-QCMD). This technique allows for the dynamic characterization of IIMP interactions with lipid bilayers without the need for purification or labeling. The method successfully reconstitutes known IIMP-membrane interactions, including the behavior of E. coli division proteins MinCDE. Additionally, it was applied to the Zorya anti-phage system, revealing specific interactions of the proteins ZorA and ZorB with bacterial membranes, while ZorE exhibited free diffusion. Overall, TXTL-QCMD demonstrates significant potential for exploring the diverse roles of IIMPs in biological systems.

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