{"external_id":{"pmid":["27821763"]},"publisher":"Proceedings of the National Academy of Sciences","publication_status":"published","date_created":"2023-09-06T12:53:48Z","month":"10","issue":"47","publication":"PNAS","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","type":"journal_article","date_updated":"2023-11-07T11:53:06Z","doi":"10.1073/pnas.1612720113","oa_version":"Published Version","extern":"1","_id":"14304","article_type":"original","status":"public","year":"2016","abstract":[{"lang":"eng","text":"Despite the recent rapid progress in cryo-electron microscopy (cryo-EM), there still exist ample opportunities for improvement in sample preparation. Macromolecular complexes may disassociate or adopt nonrandom orientations against the extended air–water interface that exists for a short time before the sample is frozen. We designed a hollow support structure using 3D DNA origami to protect complexes from the detrimental effects of cryo-EM sample preparation. For a first proof-of-principle, we concentrated on the transcription factor p53, which binds to specific DNA sequences on double-stranded DNA. The support structures spontaneously form monolayers of preoriented particles in a thin film of water, and offer advantages in particle picking and sorting. By controlling the position of the binding sequence on a single helix that spans the hollow support structure, we also sought to control the orientation of individual p53 complexes. Although the latter did not yet yield the desired results, the support structures did provide partial information about the relative orientations of individual p53 complexes. We used this information to calculate a tomographic 3D reconstruction, and refined this structure to a final resolution of ∼15 Å. This structure settles an ongoing debate about the symmetry of the p53 tetramer bound to DNA."}],"page":"E7456-E7463","day":"13","language":[{"iso":"eng"}],"author":[{"full_name":"Martin, Thomas G.","last_name":"Martin","first_name":"Thomas G."},{"first_name":"Tanmay A. M.","last_name":"Bharat","full_name":"Bharat, Tanmay A. M."},{"last_name":"Joerger","first_name":"Andreas C.","full_name":"Joerger, Andreas C."},{"first_name":"Xiao-chen","last_name":"Bai","full_name":"Bai, Xiao-chen"},{"last_name":"Praetorius","first_name":"Florian M","id":"dfec9381-4341-11ee-8fd8-faa02bba7d62","full_name":"Praetorius, Florian M"},{"first_name":"Alan R.","last_name":"Fersht","full_name":"Fersht, Alan R."},{"last_name":"Dietz","first_name":"Hendrik","full_name":"Dietz, Hendrik"},{"full_name":"Scheres, Sjors H. W.","first_name":"Sjors H. W.","last_name":"Scheres"}],"publication_identifier":{"eissn":["1091-6490"],"issn":["0027-8424"]},"article_processing_charge":"No","citation":{"mla":"Martin, Thomas G., et al. “Design of a Molecular Support for Cryo-EM Structure Determination.” PNAS, vol. 113, no. 47, Proceedings of the National Academy of Sciences, 2016, pp. E7456–63, doi:10.1073/pnas.1612720113.","ista":"Martin TG, Bharat TAM, Joerger AC, Bai X, Praetorius FM, Fersht AR, Dietz H, Scheres SHW. 2016. Design of a molecular support for cryo-EM structure determination. PNAS. 113(47), E7456–E7463.","ieee":"T. G. Martin et al., “Design of a molecular support for cryo-EM structure determination,” PNAS, vol. 113, no. 47. Proceedings of the National Academy of Sciences, pp. E7456–E7463, 2016.","short":"T.G. Martin, T.A.M. Bharat, A.C. Joerger, X. Bai, F.M. Praetorius, A.R. Fersht, H. Dietz, S.H.W. Scheres, PNAS 113 (2016) E7456–E7463.","chicago":"Martin, Thomas G., Tanmay A. M. Bharat, Andreas C. Joerger, Xiao-chen Bai, Florian M Praetorius, Alan R. Fersht, Hendrik Dietz, and Sjors H. W. Scheres. “Design of a Molecular Support for Cryo-EM Structure Determination.” PNAS. Proceedings of the National Academy of Sciences, 2016. https://doi.org/10.1073/pnas.1612720113.","apa":"Martin, T. G., Bharat, T. A. M., Joerger, A. C., Bai, X., Praetorius, F. M., Fersht, A. R., … Scheres, S. H. W. (2016). Design of a molecular support for cryo-EM structure determination. PNAS. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1612720113","ama":"Martin TG, Bharat TAM, Joerger AC, et al. Design of a molecular support for cryo-EM structure determination. PNAS. 2016;113(47):E7456-E7463. doi:10.1073/pnas.1612720113"},"intvolume":" 113","title":"Design of a molecular support for cryo-EM structure determination","scopus_import":"1","pmid":1,"quality_controlled":"1","volume":113,"date_published":"2016-10-13T00:00:00Z"}