20.500.12556/RUL-164764 Coupling of spectrin repeat modules for the assembly of nanorods and presentation of protein domains 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. polypeptide nanostructures spectrin repeat dystrophin protein self-assembly protein rods biomaterials adjustable spacing filaments ions oligomers peptides and proteins supramolecular structures and assemblies biokemija biomateriali nanostrukture polipeptidi true false true Angleški jezik Slovenski jezik Članek v reviji 2024-11-11 10:26:26 2024-11-11 10:26:27 2024-11-22 14:56:03 0000-00-00 00:00:00 2024 0 0 Str. 28748–28763 iss. 42 Vol. 18 Oct. 22, 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 1936-086X 10.1021/acsnano.4c07701 214399491 mezgec-et-al-2024-coupling-of-spectrin-repeat-modules-for-the-assembly-of-nanorods-and-presentation-of-protein-domains.pdf mezgec-et-al-2024-coupling-of-spectrin-repeat-modules-for-the-assembly-of-nanorods-and-presentation-of-protein-domains.pdf 1 3B6CA0C0D8DA57714E9F3FAE6AD7360E 43b55c81ead556734842599e4826a7972dfeec5195c48bbbbb4d7f66cf8d6d1a ef00b5ed-a00e-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=195427 https://pubs.acs.org/doi/10.1021/acsnano.4c07701 1 abd90d89-a00e-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=195426 Medicinska fakulteta Fakulteta za kemijo in kemijsko tehnologijo 0 0 0