Designed allosteric protein logic

Plaper, Tjaša (author), Merljak, Estera (author), Fink, Tina (author), Satler, Tadej (author), Ljubetič, Ajasja (author), Lainšček, Duško (author), Jazbec, Vid (author), Benčina, Mojca (author), Stevanoska, Sintija (author), Džeroski, Sašo (author), Jerala, Roman (author)

The regulation of protein function by external or internal signals is one of the key features of living organisms. The ability to directly control the function of a selected protein would represent a valuable tool for regulating biological processes. Here, we present a generally applicable regulation of proteins called INSRTR, based on inserting a peptide into a loop of a target protein that retains its function. We demonstrate the versatility and robustness of coiled-coil-mediated regulation, which enables designs for either inactivation or activation of selected protein functions, and implementation of two-input logic functions with rapid response in mammalian cells. The selection of insertion positions in tested proteins was facilitated by using a predictive machine learning model. We showcase the robustness of the INSRTR strategy on proteins with diverse folds and biological functions, including enzymes, signaling mediators, DNA binders, transcriptional regulators, reporters, and antibody domains implemented as chimeric antigen receptors in T cells. Our findings highlight the potential of INSRTR as a powerful tool for precise control of protein function, advancing our understanding of biological processes and developing biotechnological and therapeutic interventions.

• Language: English
• Keywords: cell signalling, protein folding
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: 15 str.
• Numbering: Vol. 10, art. 8
• UDC: 577
• ISSN on article: 2056-5968
• DOI: 10.1038/s41421-023-00635-y
• COBISS.SI-ID: 183898371
• Publication date in RUL: 11.11.2024

Record is a part of a journal

• Title: Cell discovery
• Publisher: Springer Nature, Chinese Academy of Sciences, Center for Excellence in Molecular Cell Science
• ISSN: 2056-5968
• COBISS.SI-ID: 525007385

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: biokemija, beljakovine, proteini, DNK, receptorji, protitelesa