IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity imex curation Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review jim.wells@ucsf.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro split luciferase complementation split luciferase complementation predetermined predetermined participant predetermined Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review I am not if we have to provide both AviTag feature and biotin tag feature.. Spike-RBD and ACE2 constructs have an AviTag™ BirA biotinylation acceptor sequence, leading to enzymatic biotinylation by BirA expressing Expi293 cells. jim.wells@ucsf.edu Accepted 2020-SEP-16 AT 11:27 BST AT 11:27 BST by PPORRAS Only protein-protein interactions imex curation imex curation Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity imex curation Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review jim.wells@ucsf.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review jim.wells@ucsf.edu Accepted 2020-SEP-16 AT 11:27 BST AT 11:27 BST by PPORRAS Only protein-protein interactions imex curation imex curation Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity imex curation Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review jim.wells@ucsf.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro bio-layer interferometry bio-layer interferometry bli bli predetermined predetermined participant predetermined Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung and James A. Wells Virus - Publications including interactions involving viral proteins This entry has been curated from a pre-print and has not been peer-reviewed. If needed, the data in this entry will be amended once the manuscript has gone through peer-review Biotinylated Spike or Spike RBD protein were immobilized on the streptavidin biosensor. After blocking with biotin, purified ACE2 proteins in solution was used as the analyte Spike-RBD and ACE2 constructs have an AviTag™ BirA biotinylation acceptor sequence, leading to enzymatic biotinylation by BirA Expi293 cells. jim.wells@ucsf.edu Accepted 2020-SEP-16 AT 11:27 BST AT 11:27 BST by PPORRAS Only protein-protein interactions imex curation imex curation ace2_human Angiotensin-converting enzyme 2 ACE2 UNQ868/PRO1885 Angiotensin-converting enzyme homolog Metalloprotease MPROT15 Angiotensin-converting enzyme-related carboxypeptidase protein protein human Homo sapiens Human MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPPNQPPVSIWLIVFGVVMGVIVVGIVILIFTGIRDRKKKNKARSGENPYASIDISKGENNPGFQNTDDVQTSF mint Carboxypeptidase which converts angiotensin I to angiotensin 1-9, a peptide of unknown function, and angiotensin II to angiotensin 1-7, a vasodilator. Also able to hydrolyze apelin-13 and dynorphin-13 with high efficiency. May be an important regulator of heart function. In case of human coronaviruses SARS and HCoV-NL63 infections, serve as functional receptor for the spike glycoprotein of both coronaviruses 8EE6EB0A931550E8 spike_sars2 Spike glycoprotein S E2 Peplomer protein 2 protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT s-34 2 5 unspecified role unspecified role neutral component neutral component purified purified T4 foldon trimerization domain tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ectodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain KV PP P0DTC2:p.Arg682_Arg685delinsAla mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR A c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 1d ace2-s-52 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 2a s-ace2-30 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 2b s-ace2-31 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 2d s-ace2-32 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 2e s-ace2-33 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified ectodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P0DTC2:p.Arg682_Arg685delinsAla mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR A P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain KV PP T4 foldon trimerization domain tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Fig. 2f ACE2-Fc concentration range:100 nM to 3.125 nM s-ace2-34 1 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range smBIT tag luciferase tag luciferase tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range lgBIT luciferase tag luciferase tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role ancillary ancillary purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association Fig. 3e The Spike protein is being pulled-down by biotin tag ace2-ace2-2 1 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range smBIT tag luciferase tag luciferase tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range lgBIT luciferase tag luciferase tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role ancillary ancillary purified purified RBD domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association Supplementary Figure 4a-b ace2-ace2-3 1 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range smBIT tag luciferase tag luciferase tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range lgBIT luciferase tag luciferase tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role ancillary ancillary purified purified ectodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range T4 foldon trimerization domain tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain KV PP P0DTC2:p.Arg682_Arg685delinsAla mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR A human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association Fig. 3f s-ace2-35 3 4 unspecified role unspecified role prey prey purified purified protease domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 5 unspecified role unspecified role bait bait purified purified ectodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P0DTC2:p.Arg682_Arg685delinsAla mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR A P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain KV PP T4 foldon trimerization domain tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Supplementary Figure 3a ACE2-Fc concentration range: 100 nM to 3.125 nM at 2-fold dilutions