{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T09:16:24Z","timestamp":1783156584311,"version":"3.54.6"},"reference-count":47,"publisher":"Elsevier BV","issue":"17","license":[{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/tdm\/userlicense\/1.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/legal\/tdmrep-license"},{"start":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T00:00:00Z","timestamp":1778457600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016828","name":"Stanford Genome Technology Center","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016828","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P30CA016087"],"award-info":[{"award-number":["P30CA016087"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01CA212608"],"award-info":[{"award-number":["R01CA212608"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["Journal of Molecular Biology"],"published-print":{"date-parts":[[2026,9]]},"DOI":"10.1016\/j.jmb.2026.169860","type":"journal-article","created":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T22:42:21Z","timestamp":1778712141000},"page":"169860","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":0,"title":["Protein Engineering-Enabled Cryo-EM Investigation of Small GTPases"],"prefix":"10.1016","volume":"438","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3833-0462","authenticated-orcid":false,"given":"Zhengshan","family":"Hu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Unnatiben Rajeshbhai","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1371-4980","authenticated-orcid":false,"given":"Eliezra","family":"Glasser","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akiko","family":"Koide","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5473-4358","authenticated-orcid":false,"given":"Shohei","family":"Koide","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"78","reference":[{"key":"10.1016\/j.jmb.2026.169860_b0005","doi-asserted-by":"crossref","first-page":"R563","DOI":"10.1016\/j.cub.2005.07.010","article-title":"Signaling interplay in Ras superfamily function","volume":"15","author":"Mitin","year":"2005","journal-title":"Curr. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0010","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1152\/physrev.2001.81.1.153","article-title":"Small GTP-binding proteins","volume":"81","author":"Takai","year":"2001","journal-title":"Physiol. Rev."},{"key":"10.1016\/j.jmb.2026.169860_b0015","doi-asserted-by":"crossref","first-page":"2457","DOI":"10.1158\/0008-5472.CAN-11-2612","article-title":"A comprehensive survey of Ras mutations in cancer","volume":"72","author":"Prior","year":"2012","journal-title":"Cancer Res."},{"key":"10.1016\/j.jmb.2026.169860_b0020","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1242\/jcs.182873","article-title":"RAS isoforms and mutations in cancer at a glance","volume":"129","author":"Hobbs","year":"2016","journal-title":"J. Cell Sci."},{"key":"10.1016\/j.jmb.2026.169860_b0025","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2305494120","article-title":"Cryo-EM structure determination of small therapeutic protein targets at 3\u00a0\u00c5-resolution using a rigid imaging scaffold","volume":"120","author":"Castells-Graells","year":"2023","journal-title":"PNAS"},{"key":"10.1016\/j.jmb.2026.169860_b0030","article-title":"Disulfi de constrained Fabs overcome target size limitation for high-resolution single-particle cryo-EM","author":"Kung","year":"2024","journal-title":"BioRxiv Prepr. Serv. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0035","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1186\/1472-6750-11-57","article-title":"Cell-free synthesis of a functional G protein-coupled receptor complexed with nanometer scale bilayer discs","volume":"11","author":"Yang","year":"2011","journal-title":"BMC Biotechnol."},{"key":"10.1016\/j.jmb.2026.169860_b0040","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1016\/j.str.2012.04.010","article-title":"Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors","volume":"20","author":"Chun","year":"2012","journal-title":"Structure"},{"key":"10.1016\/j.jmb.2026.169860_b0045","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/nature07101","article-title":"Structure of a \u03b21-adrenergic G-protein-coupled receptor","volume":"454","author":"Warne","year":"2008","journal-title":"Nature"},{"key":"10.1016\/j.jmb.2026.169860_b0050","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.1038\/s41467-020-15363-0","article-title":"Synthetic antibodies against BRIL as universal fiducial marks for single\u2212particle cryoEM structure determination of membrane proteins","volume":"11","author":"Mukherjee","year":"2020","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0055","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/s41467-024-54986-5","article-title":"A fiducial-assisted strategy compatible with resolving small MFS transporter structures in multiple conformations using cryo-EM","volume":"16","author":"Xie","year":"2025","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0060","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.jmb.2017.12.012","article-title":"Locking the elbow: improved antibody fab fragments as chaperones for structure determination","volume":"430","author":"Bailey","year":"2018","journal-title":"J. Mol. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0065","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1038\/nchembio.2231","article-title":"Inhibition of RAS function through targeting an allosteric regulatory site","volume":"13","author":"Spencer-Smith","year":"2017","journal-title":"Nature Chem. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0070","doi-asserted-by":"crossref","first-page":"3489","DOI":"10.1038\/s41388-024-03186-y","article-title":"Inhibition and degradation of NRAS with a pan-NRAS monobody","volume":"43","author":"Whaby","year":"2024","journal-title":"Oncogene"},{"key":"10.1016\/j.jmb.2026.169860_b0075","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1126\/science.2406902","article-title":"Induction of chronic myelogenous leukemia in mice by the P210bcr\/abl gene of the Philadelphia chromosome","volume":"247","author":"Daley","year":"1990","journal-title":"Science"},{"key":"10.1016\/j.jmb.2026.169860_b0080","doi-asserted-by":"crossref","first-page":"4574","DOI":"10.1073\/pnas.1218173110","article-title":"NMR-based functional profiling of RASopathies and oncogenic RAS mutations","volume":"110","author":"Smith","year":"2013","journal-title":"PNAS"},{"key":"10.1016\/j.jmb.2026.169860_b0085","doi-asserted-by":"crossref","DOI":"10.1016\/j.celrep.2021.110096","article-title":"Identification of RAS mutant biomarkers for EGFR inhibitor sensitivity using a systems biochemical approach","volume":"37","author":"McFall","year":"2021","journal-title":"Cell Rep."},{"key":"10.1016\/j.jmb.2026.169860_b0090","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.jmb.2011.12.019","article-title":"Teaching an old scaffold new tricks: monobodies constructed using alternative surfaces of the FN3 scaffold","volume":"415","author":"Koide","year":"2012","journal-title":"J. Mol. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0095","doi-asserted-by":"crossref","first-page":"7257","DOI":"10.1038\/s41467-021-27596-8","article-title":"A cryo-electron microscopy support film formed by 2D crystals of hydrophobin HFBI","volume":"12","author":"Fan","year":"2021","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0100","doi-asserted-by":"crossref","first-page":"7705","DOI":"10.1038\/s41467-024-52091-1","article-title":"Small LEA proteins mitigate air-water interface damage to fragile cryo-EM samples during plunge freezing","volume":"15","author":"Abe","year":"2024","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0105","doi-asserted-by":"crossref","first-page":"3789","DOI":"10.1038\/s41467-024-48045-2","article-title":"A conserved N-terminal motif of CUL3 contributes to assembly and E3 ligase activity of CRL3KLHL22","volume":"15","author":"Wang","year":"2024","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0110","doi-asserted-by":"crossref","first-page":"1424","DOI":"10.1016\/j.molcel.2022.02.004","article-title":"How the ends signal the end: Regulation by E3 ubiquitin ligases recognizing protein termini","volume":"82","author":"Sherpa","year":"2022","journal-title":"Mol. Cell"},{"key":"10.1016\/j.jmb.2026.169860_b0115","doi-asserted-by":"crossref","first-page":"1397","DOI":"10.1126\/science.8128219","article-title":"Crystal structure of human protein tyrosine phosphatase 1B","volume":"263","author":"Barford","year":"1994","journal-title":"Science"},{"key":"10.1016\/j.jmb.2026.169860_b0120","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1002\/pro.3148","article-title":"Monobodies and other synthetic binding proteins for expanding protein science","volume":"26","author":"Sha","year":"2017","journal-title":"Protein Sci. Publ. Protein Soc."},{"key":"10.1016\/j.jmb.2026.169860_b0125","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1016\/j.sbi.2012.06.001","article-title":"Structural insights for engineering binding proteins based on non-antibody scaffolds","volume":"22","author":"Gilbreth","year":"2012","journal-title":"Curr. Opin. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0130","doi-asserted-by":"crossref","first-page":"2177","DOI":"10.1006\/jmbi.1998.2439","article-title":"The atomic structure of protein-protein recognition sites","volume":"285","author":"Conte","year":"1999","journal-title":"J. Mol. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0135","doi-asserted-by":"crossref","first-page":"3899","DOI":"10.1038\/s41467-025-59114-5","article-title":"Inactive structures of the vasopressin V2 receptor reveal distinct binding modes for Tolvaptan and Mambaquaretin toxin","volume":"16","author":"Fouillen","year":"2025","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0140","doi-asserted-by":"crossref","first-page":"8944","DOI":"10.1038\/s41467-025-64002-z","article-title":"Structural insights into the mechanism of activation and inhibition of the prostaglandin D2 receptor 1","volume":"16","author":"Davoudinasab","year":"2025","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0145","doi-asserted-by":"crossref","first-page":"5938","DOI":"10.1038\/s41467-023-41646-3","article-title":"An inverse agonist of orphan receptor GPR61 acts by a G protein-competitive allosteric mechanism","volume":"14","author":"Lees","year":"2023","journal-title":"Nature Commun."},{"key":"10.1016\/j.jmb.2026.169860_b0150","doi-asserted-by":"crossref","unstructured":"A.W. Kahsai et al., Small-molecule modulation of \u03b2-arrestins, 2024. Preprint at https:\/\/doi.org\/10.1101\/2024.12.27.630464.","DOI":"10.1101\/2024.12.27.630464"},{"key":"10.1016\/j.jmb.2026.169860_b0155","doi-asserted-by":"crossref","first-page":"36800","DOI":"10.1038\/s41598-025-20608-3","article-title":"Structural determination of small proteins by cryo-EM using a coiled coil module strategy","volume":"15","author":"Camille","year":"2025","journal-title":"Sci. Rep."},{"key":"10.1016\/j.jmb.2026.169860_b0160","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1038\/s41592-020-01001-6","article-title":"Megabodies expand the nanobody toolkit for protein structure determination by single-particle cryo-EM","volume":"18","author":"Ucha\u0144ski","year":"2021","journal-title":"Nature Methods"},{"key":"10.1016\/j.jmb.2026.169860_b0165","doi-asserted-by":"crossref","first-page":"14924","DOI":"10.1073\/pnas.1303640110","article-title":"Dissection of the BCR-ABL signaling network using highly specific monobody inhibitors to the SHP2 SH2 domains","volume":"110","author":"Sha","year":"2013","journal-title":"PNAS"},{"key":"10.1016\/j.jmb.2026.169860_b0170","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1038\/s41589-024-01751-w","article-title":"Selective targeting of oncogenic hotspot mutations of the HER2 extracellular domain","volume":"21","author":"Bang","year":"2025","journal-title":"Nature Chem. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0175","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.jsb.2005.03.010","article-title":"Automated molecular microscopy: the new Leginon system","volume":"151","author":"Suloway","year":"2005","journal-title":"J. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0180","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1002\/pro.3967","article-title":"Leginon: new features and applications","volume":"30","author":"Cheng","year":"2021","journal-title":"Protein Sci. Publ. Protein Soc."},{"key":"10.1016\/j.jmb.2026.169860_b0185","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.jsb.2018.06.007","article-title":"Routine determination of ice thickness for cryo-EM grids","volume":"204","author":"Rice","year":"2018","journal-title":"J. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0190","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1038\/nmeth.4193","article-title":"MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy","volume":"14","author":"Zheng","year":"2017","journal-title":"Nature Methods"},{"key":"10.1016\/j.jmb.2026.169860_b0195","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.jsb.2015.08.008","article-title":"CTFFIND4: fast and accurate defocus estimation from electron micrographs","volume":"192","author":"Rohou","year":"2015","journal-title":"J. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0200","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.jsb.2009.01.002","article-title":"Appion: an integrated, database-driven pipeline to facilitate EM image processing","volume":"166","author":"Lander","year":"2009","journal-title":"J. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0205","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/nmeth.4169","article-title":"cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination","volume":"14","author":"Punjani","year":"2017","journal-title":"Nature Methods"},{"key":"10.1016\/j.jmb.2026.169860_b0210","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1038\/s41592-019-0575-8","article-title":"Positive-unlabeled convolutional neural networks for particle picking in cryo-electron micrographs","volume":"16","author":"Bepler","year":"2019","journal-title":"Nature Methods"},{"key":"10.1016\/j.jmb.2026.169860_b0215","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1038\/s41592-020-00990-8","article-title":"Non-uniform refinement: adaptive regularization improves single-particle cryo-EM reconstruction","volume":"17","author":"Punjani","year":"2020","journal-title":"Nature Methods"},{"key":"10.1016\/j.jmb.2026.169860_b0220","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera\u2013a visualization system for exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."},{"key":"10.1016\/j.jmb.2026.169860_b0225","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1107\/S2059798319011471","article-title":"Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix","volume":"75","author":"Liebschner","year":"2019","journal-title":"Acta Crystallogr. sect. Struct. Biol."},{"key":"10.1016\/j.jmb.2026.169860_b0230","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and development of Coot","volume":"66","author":"Emsley","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"10.1016\/j.jmb.2026.169860_b0235","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/pro.3943","article-title":"UCSF ChimeraX: Structure visualization for researchers, educators, and developers","volume":"30","author":"Pettersen","year":"2021","journal-title":"Protein Sci. Publ. Protein Soc."}],"container-title":["Journal of Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0022283626002330?httpAccept=text\/xml","content-type":"text\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0022283626002330?httpAccept=text\/plain","content-type":"text\/plain","content-version":"vor","intended-application":"text-mining"}],"deposited":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T08:30:10Z","timestamp":1783153810000},"score":1,"resource":{"primary":{"URL":"https:\/\/linkinghub.elsevier.com\/retrieve\/pii\/S0022283626002330"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9]]},"references-count":47,"journal-issue":{"issue":"17","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["S0022283626002330"],"URL":"https:\/\/doi.org\/10.1016\/j.jmb.2026.169860","relation":{},"ISSN":["0022-2836"],"issn-type":[{"value":"0022-2836","type":"print"}],"subject":[],"published":{"date-parts":[[2026,9]]},"assertion":[{"value":"Elsevier","name":"publisher","label":"This article is maintained by"},{"value":"Protein Engineering-Enabled Cryo-EM Investigation of Small GTPases","name":"articletitle","label":"Article Title"},{"value":"Journal of Molecular Biology","name":"journaltitle","label":"Journal Title"},{"value":"https:\/\/doi.org\/10.1016\/j.jmb.2026.169860","name":"articlelink","label":"CrossRef DOI link to publisher maintained version"},{"value":"article","name":"content_type","label":"Content Type"},{"value":"\u00a9 2026 The Author(s). Published by Elsevier Ltd.","name":"copyright","label":"Copyright"}],"article-number":"169860"}}