{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T11:12:36Z","timestamp":1773313956746,"version":"3.50.1"},"posted":{"date-parts":[[2024,3,8]]},"group-title":"Genetics","reference-count":100,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,3,8]],"date-time":"2024-03-08T00:00:00Z","timestamp":1709856000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000055","name":"NIDCD","doi-asserted-by":"crossref","award":["DC020764"],"award-info":[{"award-number":["DC020764"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000065","name":"NINDS","doi-asserted-by":"crossref","award":["NS091439"],"award-info":[{"award-number":["NS091439"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,3,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>The SARS-CoV-2 betacoronavirus infects humans through binding the Angiotensin Converting Enzyme 2 (ACE2) protein that lines the nasal cavity and lungs, followed by import into a cell utilizing the Transmembrane Protease, Serine 2 (TMPRSS2) cofactor. ACE2 binding is mediated by an approximately 200-residue portion of the SARS-CoV-2 extracellular spike protein, the receptor binding domain (RBD). Robust interactions are shown using a novel cell-based assay between an RBD membrane tethered-GFP fusion protein and a membrane-bound ACE2-Cherry fusion protein. Several observations were not predicted, including rapid and sustained interactions leading to internalization of the RBD fusion protein into ACE2-expressing cells and rapid downregulation of ACE2-Cherry fluorescence, suggesting that a membrane-associated form of RBD found on the viral coat may have long-term system-wide consequences on ACE2-expressing cells. Targeted mutation in the RBD disulfide Loop 4 led to a loss of internalization for several variants tested. However, a secreted RBD did not cause ACE2 downregulation of ACE2-Cherry fluorescence. Omicron BA.1 and BA.2 variants have altered their dependency on the amino terminus (Nt) of the ACE2 protein. In contrast, the H-CoV-NL63 RBD is only dependent on the ACE2 internal region for binding, leading to the conclusion that the RBD binding surface of ACE2 appears relatively fluid and amenable to internalizing a range of novel variants.<\/jats:p>","DOI":"10.1101\/2024.03.07.583884","type":"posted-content","created":{"date-parts":[[2024,3,8]],"date-time":"2024-03-08T23:05:19Z","timestamp":1709939119000},"source":"Crossref","is-referenced-by-count":1,"title":["Visualizing the Rapid Degradation and Differential Binding of Membrane-Bound Human ACE2 Protein Upon Binding and Internalization of the SARS-CoV-2 Spike RBD Protein"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4924-1732","authenticated-orcid":false,"given":"Paul","family":"Feinstein","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Hunter College, City University of New York, 695 Park Avenue, New York, NY 10065"},{"name":"The Graduate Center Programs in Biochemistry, Biology and CUNY Neuroscience Collaborative, 365 5th Ave, New York, NY 10016"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026031109501376000_2024.03.07.583884v2.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.03.045"},{"key":"2026031109501376000_2024.03.07.583884v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/B978-1-4557-4801-3.00157-0"},{"key":"2026031109501376000_2024.03.07.583884v2.3","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2021.635245"},{"key":"2026031109501376000_2024.03.07.583884v2.4","doi-asserted-by":"publisher","DOI":"10.7150\/ijbs.59233"},{"key":"2026031109501376000_2024.03.07.583884v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.drup.2021.100794"},{"key":"2026031109501376000_2024.03.07.583884v2.6","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-022-01780-9"},{"key":"2026031109501376000_2024.03.07.583884v2.7","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04661-w"},{"key":"2026031109501376000_2024.03.07.583884v2.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.02.052"},{"key":"2026031109501376000_2024.03.07.583884v2.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2006.01.029"},{"key":"2026031109501376000_2024.03.07.583884v2.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2023.114367"},{"key":"2026031109501376000_2024.03.07.583884v2.11","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-016-0479-5"},{"key":"2026031109501376000_2024.03.07.583884v2.12","doi-asserted-by":"crossref","unstructured":"Leon Fodoulian JT , Daniel Rossier , Madlaina Boillat , Chenda Kan , V\u00e9ronique Pauli , Kristof Egervari , Johannes A. Lobrinus , Basile N. Landis , Alan Carleton , Ivan Rodriguez . SARS-CoV-2 receptor and entry genes are expressed by sustentacular cells in the human olfactory neuroepithelium. bioRxiv 20200331013268; doi: https:\/\/doiorg\/101101\/20200331013268. 2020.","DOI":"10.1101\/2020.03.31.013268"},{"key":"2026031109501376000_2024.03.07.583884v2.13","doi-asserted-by":"crossref","unstructured":"David H. Brann TT , Caleb Weinreb , Marcela Lipovsek , Koen Van den Berge , Boying Gong , Rebecca Chance , Iain C. Macaulay , Hsin-jung Chou , Russell Fletcher , Diya Das , Kelly Street , Hector Roux de Bezieux , Yoon-Gi Choi , Davide Risso , Sandrine Dudoit , Elizabeth Purdom , Jonathan S. Mill , Ralph Abi Hachem , Hiroaki Matsunami , Darren W. Logan , Bradley J. Goldstein , Matthew S. Grubb , John Ngai , Sandeep Robert Datta . Non-neuronal expression of SARS-CoV-2 entry genes in the olfactory system suggests mechanisms underlying COVID-19-associated anosmia. bioRxiv 20200325009084; doi: https:\/\/doiorg\/101101\/20200325009084. 2020.","DOI":"10.1126\/sciadv.abc5801"},{"key":"2026031109501376000_2024.03.07.583884v2.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-33911-8"},{"key":"2026031109501376000_2024.03.07.583884v2.15","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.27845"},{"key":"2026031109501376000_2024.03.07.583884v2.16","doi-asserted-by":"publisher","DOI":"10.15252\/EMBJ.2021107821"},{"key":"2026031109501376000_2024.03.07.583884v2.17","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abc5801"},{"key":"2026031109501376000_2024.03.07.583884v2.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2020.101839"},{"key":"2026031109501376000_2024.03.07.583884v2.19","doi-asserted-by":"publisher","DOI":"10.1016\/J.CELL.2022.01.024\/ATTACHMENT\/A078E479-C54C-418F-9475-F789CFE2E8C3\/MMC2.XLSX"},{"key":"2026031109501376000_2024.03.07.583884v2.20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0074941"},{"key":"2026031109501376000_2024.03.07.583884v2.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0141712"},{"key":"2026031109501376000_2024.03.07.583884v2.22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0141696"},{"key":"2026031109501376000_2024.03.07.583884v2.23","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.C300520200"},{"key":"2026031109501376000_2024.03.07.583884v2.24","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M311191200"},{"key":"2026031109501376000_2024.03.07.583884v2.25","doi-asserted-by":"publisher","DOI":"10.1126\/science.abc0870"},{"key":"2026031109501376000_2024.03.07.583884v2.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.05.031"},{"key":"2026031109501376000_2024.03.07.583884v2.27","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0908837106"},{"key":"2026031109501376000_2024.03.07.583884v2.28","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2180-5"},{"key":"2026031109501376000_2024.03.07.583884v2.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2021.167357"},{"key":"2026031109501376000_2024.03.07.583884v2.30","doi-asserted-by":"publisher","DOI":"10.3390\/molecules27092903"},{"key":"2026031109501376000_2024.03.07.583884v2.31","doi-asserted-by":"publisher","DOI":"10.1097\/00005537-200004000-00021"},{"key":"2026031109501376000_2024.03.07.583884v2.32","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E03-05-0293"},{"key":"2026031109501376000_2024.03.07.583884v2.33","doi-asserted-by":"publisher","DOI":"10.3109\/10799899909036662"},{"key":"2026031109501376000_2024.03.07.583884v2.34","unstructured":"Parvanova I . Getting at the Surface: A Promoter and Coding Sequence Characterization of an Odorant Receptor. CUNY Academic Works. 2019."},{"key":"2026031109501376000_2024.03.07.583884v2.35","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2273.1996.tb01741.x"},{"key":"2026031109501376000_2024.03.07.583884v2.36","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E22-02-0045"},{"key":"2026031109501376000_2024.03.07.583884v2.37","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-021-01515-1"},{"key":"2026031109501376000_2024.03.07.583884v2.38","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-021-00418-x"},{"key":"2026031109501376000_2024.03.07.583884v2.39","doi-asserted-by":"publisher","DOI":"10.3390\/cells12182312"},{"key":"2026031109501376000_2024.03.07.583884v2.40","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100306"},{"key":"2026031109501376000_2024.03.07.583884v2.41","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.08.012"},{"key":"2026031109501376000_2024.03.07.583884v2.42","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.1c00560"},{"key":"2026031109501376000_2024.03.07.583884v2.43","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-021-00573-0"},{"key":"2026031109501376000_2024.03.07.583884v2.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2007.04.035"},{"key":"2026031109501376000_2024.03.07.583884v2.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.06.008"},{"key":"2026031109501376000_2024.03.07.583884v2.46","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04532-4"},{"key":"2026031109501376000_2024.03.07.583884v2.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2011.06.029"},{"key":"2026031109501376000_2024.03.07.583884v2.48","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02274-10"},{"key":"2026031109501376000_2024.03.07.583884v2.49","doi-asserted-by":"publisher","DOI":"10.1038\/srep39805"},{"key":"2026031109501376000_2024.03.07.583884v2.50","doi-asserted-by":"publisher","DOI":"10.1096\/fj.202100560R"},{"key":"2026031109501376000_2024.03.07.583884v2.51","doi-asserted-by":"publisher","DOI":"10.1021\/ja211931f"},{"key":"2026031109501376000_2024.03.07.583884v2.52","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2010146117"},{"key":"2026031109501376000_2024.03.07.583884v2.53","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009501"},{"key":"2026031109501376000_2024.03.07.583884v2.54","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3001016"},{"key":"2026031109501376000_2024.03.07.583884v2.55","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01283-20"},{"key":"2026031109501376000_2024.03.07.583884v2.56","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.10.4554"},{"key":"2026031109501376000_2024.03.07.583884v2.57","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.29349"},{"key":"2026031109501376000_2024.03.07.583884v2.58","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(92)90442-F"},{"key":"2026031109501376000_2024.03.07.583884v2.59","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.10.048"},{"key":"2026031109501376000_2024.03.07.583884v2.60","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.2008\/003962-0"},{"key":"2026031109501376000_2024.03.07.583884v2.61","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22052268"},{"key":"2026031109501376000_2024.03.07.583884v2.62","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abf1738"},{"key":"2026031109501376000_2024.03.07.583884v2.63","doi-asserted-by":"publisher","DOI":"10.1016\/j.tins.2022.11.003"},{"key":"2026031109501376000_2024.03.07.583884v2.64","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00128-22"},{"key":"2026031109501376000_2024.03.07.583884v2.65","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009246"},{"key":"2026031109501376000_2024.03.07.583884v2.66","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-021-00908-w"},{"key":"2026031109501376000_2024.03.07.583884v2.67","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2111199119"},{"key":"2026031109501376000_2024.03.07.583884v2.68","doi-asserted-by":"publisher","DOI":"10.1002\/prot.26024"},{"key":"2026031109501376000_2024.03.07.583884v2.69","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3001989"},{"key":"2026031109501376000_2024.03.07.583884v2.70","doi-asserted-by":"publisher","DOI":"10.1016\/j.dsx.2020.04.020"},{"key":"2026031109501376000_2024.03.07.583884v2.71","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-28961-x"},{"key":"2026031109501376000_2024.03.07.583884v2.72","doi-asserted-by":"publisher","DOI":"10.1002\/advs.201903204"},{"key":"2026031109501376000_2024.03.07.583884v2.73","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2025.1517585"},{"key":"2026031109501376000_2024.03.07.583884v2.74","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-75192-9"},{"key":"2026031109501376000_2024.03.07.583884v2.75","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMc2209651"},{"key":"2026031109501376000_2024.03.07.583884v2.76","doi-asserted-by":"crossref","unstructured":"Zhu J , Su, Y. , and Tang, Y. Disrupting ACE2 Dimerization Mitigates the Infection by SARS-CoV-2 Pseudovirus. Frontiers in Virology. 2022;Virol., 03 July 2022(2).","DOI":"10.3389\/fviro.2022.916700"},{"key":"2026031109501376000_2024.03.07.583884v2.77","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.101175"},{"key":"2026031109501376000_2024.03.07.583884v2.78","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2023.04.088"},{"key":"2026031109501376000_2024.03.07.583884v2.79","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-020-0458-z"},{"key":"2026031109501376000_2024.03.07.583884v2.80","doi-asserted-by":"publisher","DOI":"10.1016\/j.peptides.2022.170898"},{"key":"2026031109501376000_2024.03.07.583884v2.81","doi-asserted-by":"publisher","DOI":"10.1126\/science.abd9909"},{"key":"2026031109501376000_2024.03.07.583884v2.82","doi-asserted-by":"publisher","DOI":"10.1016\/j.csbj.2021.06.041"},{"key":"2026031109501376000_2024.03.07.583884v2.83","doi-asserted-by":"publisher","DOI":"10.1007\/s00284-022-02807-7"},{"key":"2026031109501376000_2024.03.07.583884v2.84","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-022-07700-x"},{"key":"2026031109501376000_2024.03.07.583884v2.85","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacbts.2020.10.003"},{"key":"2026031109501376000_2024.03.07.583884v2.86","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.765300"},{"key":"2026031109501376000_2024.03.07.583884v2.87","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.abf1117"},{"key":"2026031109501376000_2024.03.07.583884v2.88","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.61390"},{"key":"2026031109501376000_2024.03.07.583884v2.89","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.658428"},{"key":"2026031109501376000_2024.03.07.583884v2.90","doi-asserted-by":"publisher","DOI":"10.3389\/fddsv.2021.789710"},{"key":"2026031109501376000_2024.03.07.583884v2.91","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmed.2022.04.007"},{"key":"2026031109501376000_2024.03.07.583884v2.92","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejim.2021.04.019"},{"key":"2026031109501376000_2024.03.07.583884v2.93","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejim.2021.06.013"},{"key":"2026031109501376000_2024.03.07.583884v2.94","doi-asserted-by":"publisher","DOI":"10.1038\/s44161-023-00222-0"},{"key":"2026031109501376000_2024.03.07.583884v2.95","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2027906"},{"key":"2026031109501376000_2024.03.07.583884v2.96","doi-asserted-by":"publisher","DOI":"10.1001\/jamanetworkopen.2021.43955"},{"key":"2026031109501376000_2024.03.07.583884v2.97","doi-asserted-by":"publisher","DOI":"10.1006\/mcne.2002.1106"},{"key":"2026031109501376000_2024.03.07.583884v2.98","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b07461"},{"key":"2026031109501376000_2024.03.07.583884v2.99","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.0c02125"},{"key":"2026031109501376000_2024.03.07.583884v2.100","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-78711-6"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.03.07.583884","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T16:50:26Z","timestamp":1773247826000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.03.07.583884"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,8]]},"references-count":100,"URL":"https:\/\/doi.org\/10.1101\/2024.03.07.583884","relation":{},"subject":[],"published":{"date-parts":[[2024,3,8]]},"subtype":"preprint"}}