{"doi":"10.1101/2020.07.27.221333","title":"Hierarchical design of multi-scale protein complexes by combinatorial assembly of oligomeric helical bundle and repeat protein building blocks","abstract":"Abstract A goal of de novo protein design is to develop a systematic and robust approach to generating complex nanomaterials from stable building blocks. Due to their structural regularity and simplicity, a wide range of monomeric repeat proteins and oligomeric helical bundle structures have been designed and characterized. Here we describe a stepwise hierarchical approach to building up multi-component symmetric protein assemblies using these structures. We first connect designed helical repeat proteins (DHRs) to designed helical bundle proteins (HBs) to generate a large library of heterodimeric and homooligomeric building blocks; the latter have cyclic symmetries ranging from C2 to C6. All of the building blocks have repeat proteins with accessible termini, which we take advantage of in a second round of architecture guided rigid helical fusion (WORMS) to generate larger symmetric assemblies including C3 and C5 cyclic and D2 dihedral rings, a tetrahedral cage, and a 120 subunit icosahedral cage. Characterization of the structures by small angle x-ray scattering, x-ray crystallography, and cryo-electron microscopy demonstrates that the hierarchical design approach can accurately and robustly generate a wide range of macromolecular assemblies; with a diameter of 43nm, the icosahedral nanocage is the largest structurally validated designed cage to date. The computational methods and building block sets described here provide a very general route to new de novo designed symmetric protein nanomaterials.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":121081,"datarank":0.32958368660043297,"base_score":2.1972245773362196,"endowment":2.1972245773362196,"self_citation_contribution":0.32958368660043297,"citation_network_contribution":0.0,"self_endowment_contribution":0.32958368660043297,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9531,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":560137,"name":"Rubul Mout","orcid":"0000-0001-6125-5873","position":1,"is_corresponding":false},{"id":249505,"name":"William Sheffler","orcid":"0000-0002-3659-9559","position":2,"is_corresponding":false},{"id":560138,"name":"Natasha I. 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Bera","orcid":"0000-0001-9473-2912","position":8,"is_corresponding":false},{"id":557871,"name":"Alexis Courbet","orcid":"0000-0003-0539-7011","position":9,"is_corresponding":false},{"id":268184,"name":"Alex Kang","orcid":"0000-0001-5487-0499","position":10,"is_corresponding":false},{"id":353804,"name":"TJ Brunette","orcid":"0000-0003-0748-8224","position":11,"is_corresponding":false},{"id":560139,"name":"Una Nattermann","orcid":"0000-0001-7837-7113","position":12,"is_corresponding":false},{"id":561070,"name":"Evelyn Tsai","orcid":null,"position":13,"is_corresponding":false},{"id":561071,"name":"Ayesha Saleem","orcid":null,"position":14,"is_corresponding":false},{"id":225386,"name":"Cameron M. Chow","orcid":"0000-0001-5351-6412","position":15,"is_corresponding":false},{"id":300956,"name":"Damian C. 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