Segmentation strategy of de novo designed four-helical bundles expands protein oligomerization modalities for cell regulation

Merljak, Estera (author), Malovrh, Benjamin (author), Jerala, Roman (author)

Protein–protein interactions govern most biological processes. New protein assemblies can be introduced through the fusion of selected proteins with di/oligomerization domains, which interact specifically with their partners but not with other cellular proteins. While four-helical bundle proteins (4HB) have typically been assembled from two segments, each comprising two helices, here we show that they can be efficiently segmented in various ways, expanding the number of combinations generated from a single 4HB. We implement a segmentation strategy of 4HB to design two-, three-, or four-chain combinations for the recruitment of multiple protein components. Different segmentations provide new insight into the role of individual helices for 4HB assembly. We evaluate 4HB segmentations for potential use in mammalian cells for the reconstitution of a protein reporter, transcriptional activation, and inducible 4HB assembly. Furthermore, the implementation of trimerization is demonstrated as a modular chimeric antigen receptor for the recognition of multiple cancer antigens.

• Language: English
• Keywords: molecular engineering, synthetic biology
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: 12 str.
• Numbering: Vol. 14, art. 1995
• PID: 20.500.12556/RUL-155351
• UDC: 577
• ISSN on article: 2041-1723
• DOI: 10.1038/s41467-023-37765-6
• COBISS.SI-ID: 150022147
• Publication date in RUL: 27.03.2024

Record is a part of a journal

• Title: Nature communications
• Shortened title: Nat. commun.
• Publisher: Springer Nature
• ISSN: 2041-1723
• COBISS.SI-ID: 2315876

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, proteini, beljakovine, antigenski receptorji, rakava obolenja

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: P4-0176
• Name: Molekularna biotehnologija: od dinamike bioloških sistemov do aplikacij