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/ A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. Nature structural & molecular biology 2021 imex curation Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) see pmid 33727252: which refers to this one-from same group. The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu 24-May-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro 3d-em 3D electron microscopy 3D-EM electron tomog structure determination identification by structure determination identification by structure determination structure determination Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu Accepted 2021-APR-05 AT 10:57 BST AT 10:57 BST by KALPANAP Nature structural & molecular biology 2021 Only protein-protein interactions imex curation imex curation A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. Nature structural & molecular biology 2021 imex curation Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) see pmid 33727252: which refers to this one-from same group. The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu 24-May-2021: 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 Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu Accepted 2021-APR-05 AT 10:57 BST AT 10:57 BST by KALPANAP Nature structural & molecular biology 2021 Only protein-protein interactions imex curation imex curation A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent. Nature structural & molecular biology 2021 imex curation Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) see pmid 33727252: which refers to this one-from same group. The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu 24-May-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro cleavage cleavage assay cleavage predetermined predetermined participant predetermined Xiao T., Lu J., Zhang J., Johnson RI., McKay LGA., Storm N., Lavine CL., Peng H., Cai Y., Rits-Volloch S., Lu S., Quinlan BD., Farzan M., Seaman MS., Griffiths A., Chen B. Virus - Publications including interactions involving viral proteins Mol. Sieving Extended Data figure 1 not added - supposed to show "monomeric soluble ACE2 (residues 18–615) in complex with a stabilized soluble SARS-CoV-2 S protein trimer";the M. wt on the SDS-PAGE gel does not show this (ACE2 tagless monomer construct =69kDa, Spike tagless construct monomer =134kDa.) FRET based assay:On cleavage of the peptide by the protease, the quencher (DNP) and fluorophore (MCA) are separated giving a large increase in fluorescence. The Specific Activity was calculated as the amount of the released fluorophore divided by the reaction time and the amount of the ACE2 protein using the data within the initial linear phase. The authors have stated that the "m" in ACE2m-615-Fc stands for "mutated", with mutations in the peptidase active site. Peptidase activity assay; BioVision, the suppliers of the synthetic peptide substrate from the ACE2 activity kit have confirmed the sequence of the substrate was MCA-APK-(Dnp)-OH (Mca-Ala-Pro-Lys(Dnp)-OH) as in Catalog #: B2726. bchen@crystal.harvard.edu Accepted 2021-APR-05 AT 10:57 BST AT 10:57 BST by KALPANAP Nature structural & molecular biology 2021 Only protein-protein interactions imex curation imex curation 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 mca-apk-dnp-oh Mca-APK(Dnp)-OH peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) APK yes 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 ace2-s-86 1 6 unspecified role unspecified role 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 13:24:21 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. direct interaction direct interaction Table 1 ace2-s-87 1 6 unspecified role unspecified role 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 11:04:46 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. direct interaction direct interaction Table 1 ace2-s-88 1 6 unspecified role unspecified role 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 12:05:40 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. direct interaction direct interaction Table 1 ace2-s-89 1 4 unspecified role unspecified role neutral component neutral component purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 13:13:34 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. direct interaction direct interaction Table 1 ace2-s-90 2 6 unspecified role unspecified role prey prey 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain 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. 4 unspecified role unspecified role bait bait purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 13:33:34 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction 2B, Table 2+3. Curves for monomeric ACE2 fitted a 1:1 binding model. I am assuming the stated 1:1 binding means one mol binds to one mol? - so 3 mols to 3. ACE2-615-a monomer. s-ace2-75 2 6 unspecified role unspecified role prey prey purified purified 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 ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. 4 unspecified role unspecified role bait bait purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 12:04:20 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction 2B, Table 2+3. ACE2-740-Fc - a dimer. Curves for oligomeric ACE2 variants fitted a bivalent binding model. The bivalent model describes the binding of a bivalent analyte to immobilized ligand, where one analyte molecule can bind to one or two ligand molecules. As a result of binding of one analyte molecule to two ligand sites, the overall binding is strengthened compared with 1:1 binding. s-ace2-76 2 6 unspecified role unspecified role prey prey 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain T4 phage fibritin trimerization motif and slightly longer linker (13 residues) tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9BYF1:p.Thr27Tyr mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain T Y Thu Apr 22 11:45:07 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr27Tyr' T4 phage fibritin trimerization motif with linker (11 residues) tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9BYF1:p.Thr27Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain T W Thu Apr 22 12:33:41 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr27Trp' Q9BYF1:p.His34Trp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain H W Thu Apr 22 13:03:56 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His34Trp' Q9BYF1:p.Lys353Tyr mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain K Y Thu Apr 22 11:43:56 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys353Tyr' 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. 4 unspecified role unspecified role bait bait purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 11:56:53 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction 2B, Table 2+3, Extended data 7+8. ACE2-615-foldon- a trimer. Curves for oligomeric ACE2 variants fitted a bivalent binding model. The bivalent model describes the binding of a bivalent analyte to immobilized ligand, where one analyte molecule can bind to one or two ligand molecules. As a result of binding of one analyte molecule to two ligand sites, the overall binding is strengthened compared with 1:1 binding. A] I do not see any sequence for linkers; I have entered the "LL-foldon" as my "slightly longer linker-13 residues", as a separate "tag" rather than as a mutation increasing strength. If entered as a mutation with as "c-c" for range I am not sure if this is allowed? (might break something?) Strictly speaking it should NOT be entered as a "tag" as it does affect the interaction;but not sure if/how this could be done differently. (any ideas??) B] According to the bottom construct in figure 2A there is NO C-terminal HIS-Tag - however from reading the text + Methods (purification), I think this is unlikely, so have assumed it is a typo? -and left the HIS tag in for the mutants. s-ace2-77 2 6 unspecified role unspecified role prey prey purified purified 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain Q9BYF1:p.[His374Asn;His378Asn] mutation mutation certain certain sequence position certain certain certain sequence position certain H N certain certain sequence position certain certain certain sequence position certain H N Thu Apr 22 10:41:54 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[His374Asn;His378Asn]' 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. 4 unspecified role unspecified role bait bait purified purified P0DTC2:p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain RRAR GGSG certain certain sequence position certain certain certain sequence position certain KV PP Thu Apr 22 11:56:15 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg682_Arg685delinsGlyGlySerGly;Lys986_Val987delinsProPro]]' 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 phage fibritin trimerization motif tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Table 2+3. Extended Data 8. ACE2m-615-Fc -a dimer. Curves for oligomeric ACE2 variants fitted a bivalent binding model. The bivalent model describes the binding of a bivalent analyte to immobilized ligand, where one analyte molecule can bind to one or two ligand molecules. As a result of binding of one analyte molecule to two ligand sites, the overall binding is strengthened compared with 1:1 binding. ace2-mca_apk(dnp)_oh-1 3 6 enzyme enzyme 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 ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain 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. 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 3A All the trimeric forms showed essentially the same specific activity; the monomeric ACE2615 and the dimeric ACE2740-Fc had lower specific activities. ACE2-615-a monomer. Specific Activity:4.54mU/ng ace2-mca_apk(dnp)_oh-2 3 6 enzyme enzyme neutral component neutral component purified purified 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 ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 3A All the trimeric forms showed essentially the same specific activity; the monomeric ACE2615 and the dimeric ACE2740-Fc had lower specific activities. ACE2-740-Fc - a dimer. Specific Activity:3.59 mU/ng ace2-mca_apk(dnp)_oh-3 3 6 enzyme enzyme neutral component neutral component purified purified ecd sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain T4 phage fibritin trimerization motif and slightly longer linker (13 residues) tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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 Q9BYF1:p.Thr27Tyr mutation mutation certain certain sequence position certain certain certain sequence position certain T Y Thu Apr 22 13:32:40 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr27Tyr' T4 phage fibritin trimerization motif with linker (11 residues) tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9BYF1:p.Thr27Trp mutation mutation certain certain sequence position certain certain certain sequence position certain T W Thu Apr 22 13:10:18 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr27Trp' Q9BYF1:p.His34Trp mutation mutation certain certain sequence position certain certain certain sequence position certain H W Thu Apr 22 11:02:38 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His34Trp' Q9BYF1:p.Lys353Tyr mutation mutation certain certain sequence position certain certain certain sequence position certain K Y Thu Apr 22 11:44:24 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys353Tyr' 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. 5 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 3A All the trimeric forms showed essentially the same specific activity; the monomeric ACE2615 and the dimeric ACE2740-Fc had lower specific activities. ACE2-615-foldon- a trimer. Specific Activities:range 5.4 to 7.2 (K353Y)mU/ng.