Coupling of spectrin repeat modules for the assembly of nanorods and presentation of protein domains

Mezgec, Klemen (author), Snoj, Jaka (author), Ulčakar, Liza (author), Ljubetič, Ajasja (author), Tušek-Žnidarič, Magda (author), Škarabot, Miha (author), Jerala, Roman (author)

Modular protein engineering is a powerful approach for fabricating high-molecular-weight assemblies and biomaterials with nanoscale precision. Herein, we address the challenge of designing an extended nanoscale filamentous architecture inspired by the central rod domain of human dystrophin, which protects sarcolemma during muscle contraction and consists of spectrin repeats composed of three-helical bundles. A module of three tandem spectrin repeats was used as a rigid building block self-assembling via coiled-coil (CC) dimer-forming peptides. CC peptides were precisely integrated to maintain the spectrin α-helix continuity in an appropriate frame to form extended nanorods. An orthogonal set of customizable CC heterodimers was harnessed for modular rigid domain association, which could be additionally regulated by metal ions and chelators. We achieved a robust assembly of rigid rods several micrometers in length, determined by atomic force microscopy and negative stain transmission electron microscopy. Furthermore, these rigid rods can serve as a scaffold for the decoration of diverse proteins or biologically active peptides along their length with adjustable spacing up to tens of nanometers, as confirmed by the DNA-PAINT super-resolution microscopy. This demonstrates the potential of modular bottom-up protein engineering and tunable CCs for the fabrication of functionalized protein biomaterials.

• Language: English
• Keywords: polypeptide nanostructures, spectrin repeat, dystrophin, protein self-assembly, protein rods, biomaterials, adjustable spacing, filaments, ions, oligomers, peptides and proteins, supramolecular structures and assemblies
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine; FKKT - Faculty of Chemistry and Chemical Technology
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 28748–28763
• Numbering: Vol. 18, iss. 42
• PID: 20.500.12556/RUL-164764
• UDC: 577
• ISSN on article: 1936-086X
• DOI: 10.1021/acsnano.4c07701
• COBISS.SI-ID: 214399491
• Publication date in RUL: 11.11.2024

Record is a part of a journal

• Title: ACS nano
• Shortened title: ACS nano
• Publisher: American Chemical Society
• ISSN: 1936-086X
• COBISS.SI-ID: 513574937

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, biomateriali, nanostrukture, polipeptidi