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- W4310288316 abstract "Reversible membrane targeting of proteins is one of the key regulators of cellular interaction networks, for example, for signaling and polarization. So-called membrane switches are thus highly attractive targets for the design of minimal cells but have so far been tricky to reconstitute in vitro. Here, we introduce cell-free prenylated protein synthesis (CFpPS), which enables the synthesis and membrane targeting of proteins in a single reaction mix including the prenylation machinery. CFpPS can confer membrane affinity to any protein via addition of a 4-peptide motif to its C-terminus and offers robust production of prenylated proteins not only in their soluble forms but also in the direct vicinity of biomimetic membranes. Thus, CFpPS enabled us to reconstitute the prenylated polarity hub Cdc42 and its regulatory protein in vitro, implementing a key membrane switch. We propose CFpPS to be a versatile and effective platform for engineering complex features, such as polarity induction, in synthetic cells." @default.
- W4310288316 created "2022-11-30" @default.
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- W4310288316 date "2022-11-29" @default.
- W4310288316 modified "2023-09-26" @default.
- W4310288316 title "Reconstitution of a Reversible Membrane Switch via Prenylation by One-Pot Cell-Free Expression" @default.
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- W4310288316 doi "https://doi.org/10.1021/acssynbio.2c00406" @default.
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