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bioRxiv · 10.1101/2022.06.03.494683

Designed allosteric protein logic

Plaper, T.·Merljak, E.·Fink, T.·Lainscek, D.·Satler, T.·Jazbec, V.·Bencina, M.·Jerala, R.
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Abstract

Regulation of the activity of proteins enables control of complex cellular processes. Allosteric regulation has been introduced individually into few natural proteins. Here, we present a generally applicable regulation of diverse proteins called INSRTR (inserted peptide structure regulator), based on inserting a short unstructured peptide into a solvent-accessible loop that retains protein function. Function of the target protein can be inactivated by the addition of a peptide that forms a rigid coiled-coil dimer. This platform enables the construction of ON/OFF protein switches, their regulation by small molecules, and Boolean logic functions with a rapid response in mammalian cells. INSRTR can be used to regulate a wide range of proteins, as demonstrated on ten members of protein families with diverse biological activities including enzymes, signaling mediators, DNA binders/transcriptional regulators, fluorescent protein, and antibodies regulating chimeric antigen receptor. INSRTR platform presents an extraordinary potential for regulating biological systems and applications. One sentence summaryAuthors have designed a widely applicable system to activate or inactivate function of diverse proteins or form Boolean logic gates based on formation of a coiled-coil dimer within protein loops and demonstrated its implementation on a range of 10 diverse proteins.

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Plaper, T., Merljak, E., Fink, T., Lainscek, D., Satler, T., Jazbec, V., Bencina, M., Jerala, R.. 2022-06-03. Designed allosteric protein logic. https://doi.org/10.1101/2022.06.03.494683

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