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/ http://www.ebi.ac.uk/ lu-2020-1 in vitro In vitro cleavage cleavage assay cleavage predetermined predetermined participant predetermined Lu J., Sun PD. Virus - Publications including interactions involving viral proteins ACE2 was supplied from two suppliers, each using a different expression system;one is "Insect" cells, the other 293 cells. It is not stated which/both is used in these assays. The peptide substrates contain a highly fluorescent 7-methoxycoumarin group that is efficiently quenched by resonance energy transfer to the 2,4-dinitrophenyl group. Peptidases that are capable of cleaving an amide bond between the fluorescent group and the quencher group, cause an increase in fluorescence. SARS2 Spike protein found from Wrapp-2020, pmid:32075877. psun@nih.gov The Journal of biological chemistry 2020 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus lu-2020-3 in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Lu J., Sun PD. Virus - Publications including interactions involving viral proteins SARS2 Spike protein found from Wrapp-2020, pmid:32075877. psun@nih.gov The Journal of biological chemistry 2020 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus 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 peptide-mca-yvadapk-dnp 7-methoxycoumarin-YVADAPK(2,4-dinitrophenyl)-OH peptide peptide polypeptide oligopeptide oligopeptide polypeptide in vitro In vitro YVADAPK 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 peptide-mca-rppgfsafk-dnp Mca-RPPGFSAFK(Dnp)-OH peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) RPPGFSAFK yes spike_sars Spike glycoprotein S E2 Peplomer protein 2 protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus MFIFLLFLTLTSGSDLDRCTTFDDVQAPNYTQHTSSMRGVYYPDEIFRSDTLYLTQDLFLPFYSNVTGFHTINHTFGNPVIPFKDGIYFAATEKSNVVRGWVFGSTMNNKSQSVIIINNSTNVVIRACNFELCDNPFFAVSKPMGTQTHTMIFDNAFNCTFEYISDAFSLDVSEKSGNFKHLREFVFKNKDGFLYVYKGYQPIDVVRDLPSGFNTLKPIFKLPLGINITNFRAILTAFSPAQDIWGTSAAAYFVGYLKPTTFMLKYDENGTITDAVDCSQNPLAELKCSVKSFEIDKGIYQTSNFRVVPSGDVVRFPNITNLCPFGEVFNATKFPSVYAWERKKISNCVADYSVLYNSTFFSTFKCYGVSATKLNDLCFSNVYADSFVVKGDDVRQIAPGQTGVIADYNYKLPDDFMGCVLAWNTRNIDATSTGNYNYKYRYLRHGKLRPFERDISNVPFSPDGKPCTPPALNCYWPLNDYGFYTTTGIGYQPYRVVVLSFELLNAPATVCGPKLSTDLIKNQCVNFNFNGLTGTGVLTPSSKRFQPFQQFGRDVSDFTDSVRDPKTSEILDISPCSFGGVSVITPGTNASSEVAVLYQDVNCTDVSTAIHADQLTPAWRIYSTGNNVFQTQAGCLIGAEHVDTSYECDIPIGAGICASYHTVSLLRSTSQKSIVAYTMSLGADSSIAYSNNTIAIPTNFSISITTEVMPVSMAKTSVDCNMYICGDSTECANLLLQYGSFCTQLNRALSGIAAEQDRNTREVFAQVKQMYKTPTLKYFGGFNFSQILPDPLKPTKRSFIEDLLFNKVTLADAGFMKQYGECLGDINARDLICAQKFNGLTVLPPLLTDDMIAAYTAALVSGTATAGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKQIANQFNKAISQIQESLTTTSTALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQAAPHGVVFLHVTYVPSQERNFTTAPAICHEGKAYFPREGVFVFNGTSWFITQRNFFSPQIITTDNTFVSGNCDVVIGIINNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYVWLGFIAGLIAIVMVTILLCCMTSCCSCLKGACSCGSCCKFDEDDSEPVLKGVKLHYT p01042-pro_0000006688 Bradykinin KNG1 BDK KNG Alpha-2-thiol proteinase inhibitor Fitzgerald factor High molecular weight kininogen Williams-Fitzgerald-Flaujeac factor protein protein human Homo sapiens Human RPPGFSPFR 381 389 p01019-pro_0000032458 Angiotensin-2 AGT SERPINA8 Serpin A8 protein protein human Homo sapiens Human DRVYIHPF 34 41 mln-4760 MLN-4760 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ace2-peptide_mca_yvadapk_dnp-1 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 1A/C ,2,3,4, 5, Supp. fig 1, Table 1 Km: average value =36um (47uM +/-10 for 150mM NaCl and 25uM +/-10 for 300mM NaCl). Kcat/Km = 8.63 X10E2 /M/sec (7.08 X10E3 /M/sec for 150mM NaCl and 1.66 X10E4 /M/sec for 300mM NaCl). ace2-peptide_mca_yvadapk_dnp-2 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 2A,3,4,5A, Table 1 ace2-peptide_mca_rppgfsafk_dnp-1 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 2C,3,Supp. fig 3 Figure 2C/D:only SARS-CoV-2 RBD but not SARS-CoV RBD enhanced ACE2 cleavage of the BK-analog. ace2-peptide_mca_rppgfsafk_dnp-2 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 1B/D, 2,3,4, Supp. fig 1, 3, Table 1 Average Kcat/Km = 8.63 X10E2 /M/sec (7.06 X10E2 /M/sec for 150mM NaCl and 1.02 X10E3 /M/sec for 300mM NaCl). ace2-peptide_mca_yvadapk_dnp-3 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946541 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 9inse 9inse N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 2B, 3A ace2-peptide_mca_yvadapk_dnp-4 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946542 inhibitor inhibitor neutral component neutral component des-BK sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 5A, Table 1 ace2-agt-5 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946543 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 5A, Table 1 150mM NaCl ace2-kng1-3 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946542 inhibitor inhibitor neutral component neutral component des-BK sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) 3946535 N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 5B BK and desBK appeared to inhibit the cleavage of caspase-1 substrate (Mca-YVADAPK(Dnp) at a concentration of 640um, in the absence of the Spike rbd. 150mM NaCl ace2-agt-6 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946543 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 5B, table 1 In the absence of SARS-CoV-2 RBD binding, only Ang II but not BK or desBK inhibited the cleavage of caspase-1 substrate (Mca-YVADAPK(Dnp)-OH). ace2-kng1-4 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946542 inhibitor inhibitor neutral component neutral component des-BK sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false Supp. figure 3 In the absence of Spike-rbd protein the BK and des-BK peptides had no inhibitory effect. ace2-agt-7 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946543 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 Supp. figure 3 150mM NaCl ace2-agt-8 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946543 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3946535 Supp. figure 3 150mM NaCl ace2-peptide_mca_rppgfsafk_dnp-3 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946544 inhibitor inhibitor unspecified role unspecified role chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3B MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-peptide_mca_yvadapk_dnp-5 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946544 inhibitor inhibitor unspecified role unspecified role N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3A MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-peptide_mca_yvadapk_dnp-6 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946539 stimulator stimulator neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false T4 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false C-terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false HRV3C protease cleavage site tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain false P0DTC2:p.[Arg682_Arg683delinsGlySer;Arg685Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f Stoichiometry: 3.0 cleavage cleavage reaction cleavage false false false 1A/C, 3,4, Supp. Figure 1,Table 1 Km: average value =24.7um (28.2uM +/-0.6 for 150mM NaCl and 21.1uM +/-6.8 for 300mM NaCl). Kcat/Km = 2.06 X10E4 /M/sec for 150mM NaCl and 3.71 X10E4 /M/sec for 300mM NaCl). ace2-peptide_mca_rppgfsafk_dnp-4 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false T4 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false C-terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false HRV3C protease cleavage site tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain false P0DTC2:p.[Arg682_Arg683delinsGlySer;Arg685Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f Stoichiometry: 3.0 N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 1B/D, 3,4, Supp. figure 1,Table 1 Km: ~1.9uM for both 150 and 300mM [NaCl]. Kcat/Km:average value =9.5 X10E3 /M/sec (7.5 X10E3 /M/sec for 150mM NaCl and 1.14X10E4 /M/sec for 300mM NaCl) ace2-mln_4760-1 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false T4 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false C-terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false HRV3C protease cleavage site tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain false P0DTC2:p.[Arg682_Arg683delinsGlySer;Arg685Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f Stoichiometry: 3.0 N/A 3946544 inhibitor inhibitor unspecified role unspecified role chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3B MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-mln_4760-2 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. N/A 3946544 inhibitor inhibitor unspecified role unspecified role chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3B MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-peptide_mca_yvadapk_dnp-7 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946541 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 9inse 9inse N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3B ace2-peptide_mca_rppgfsafk_dnp-5 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946544 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946541 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 9inse 9inse N/A 3946540 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3B ace2-mln_4760-3 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946539 stimulator stimulator neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false T4 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false C-terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false HRV3C protease cleavage site tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain false P0DTC2:p.[Arg682_Arg683delinsGlySer;Arg685Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f Stoichiometry: 3.0 N/A 3946544 inhibitor inhibitor unspecified role unspecified role N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3A MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-mln_4760-4 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946544 inhibitor inhibitor unspecified role unspecified role N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946539 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. cleavage cleavage reaction cleavage false false false 3A MLN-4760 blocked ACE2 activity in the absence or presence of SARS-CoV-2 spike, SARS-CoV-2 RBD, or SARS-CoV RBD proteins. ace2-mln_4760-5 3946535 N/A 3946537 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3946544 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3946541 stimulator stimulator neutral component neutral component purified purified 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 false rbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 9inse 9inse N/A 3946538 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage false false false 3A ace2-s-74 3946536 N/A 3946537 unspecified role unspecified role bait bait purified purified ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 3.0 N/A 3946539 unspecified role unspecified role prey prey purified purified T4 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false C-terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false HRV3C protease cleavage site tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0DTC2:p.Lys986_Val987delinsProPro mutation mutation certain certain sequence position certain certain certain sequence position certain false P0DTC2:p.[Arg682_Arg683delinsGlySer;Arg685Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f Stoichiometry: 3.0 direct interaction direct interaction false false false 3946536 Supp. Figure 2