20.500.12556/RUL-152674 Crystal structure of de novo designed coiled-coil protein origami triangle Coiled-coil protein origami (CCPO) uses modular coiled-coil building blocks and topological principles to design polyhedral structures distinct from those of natural globular proteins. While the CCPO strategy has proven successful in designing diverse protein topologies, no high-resolution structural information has been available about these novel protein folds. Here we report the crystal structure of a single-chain CCPO in the shape of a triangle. While neither cyclization nor the addition of nanobodies enabled crystallization, it was ultimately facilitated by the inclusion of a GCN$_2$ homodimer. Triangle edges are formed by the orthogonal parallel coiled-coil dimers P1:P2, P3:P4, and GCN$_2$ connected by short linkers. A triangle has a large central cavity and is additionally stabilized by side-chain interactions between neighboring segments at each vertex. The crystal lattice is densely packed and stabilized by a large number of contacts between triangles. Interestingly, the polypeptide chain folds into a trefoil-type protein knot topology, and AlphaFold2 fails to predict the correct fold. The structure validates the modular CC-based protein design strategy, providing molecular insight underlying CCPO stabilization and new opportunities for the design. crystal structure crystals noncovalent interactions oligomers peptides proteins biokemija proteinski origami obvite vijačnice CCPO DNK true false true Angleški jezik Slovenski jezik Članek v reviji 2023-12-04 10:20:26 2023-12-04 10:20:27 2023-12-05 03:41:06 0000-00-00 00:00:00 2023 0 0 Str. 16995–17000 iss. 31 Vol. 145 Aug. 9, 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 1520-5126 10.1021/jacs.3c05531 169812995 satler-et-al-2023-crystal-structure-of-de-novo-designed-coiled-coil-protein-origami-triangle.pdf satler-et-al-2023-crystal-structure-of-de-novo-designed-coiled-coil-protein-origami-triangle.pdf 1 57AA35C357A4DA8D5CA8EB161918A4C9 c830c335bf3503862e79bcc81ec09430fb8a5455e4918b2eac892f2f3956ce43 b3cc88e2-9286-11ee-9ca9-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=178279 https://pubs.acs.org/doi/10.1021/jacs.3c05531 0 90de9e61-9286-11ee-9ca9-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=178278 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0