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/ Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. Cell 2020 imex curation Yurkovetskiy L., Wang X., Pascal KE., Tomkins-Tinch C., Nyalile TP., Wang Y., Baum A., Diehl WE., Dauphin A., Carbone C., Veinotte K., Egri SB., Schaffner SF., Lemieux JE., Munro JB., Rafique A., Barve A., Sabeti PC., Kyratsous CA., Dudkina NV., Shen K., Luban J. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus 17/12/20:- Addgene website NOW has constructs listed -but probably not the one we need. (as of 27/10/20 Addgene do not have the plasmids used in this publication visible on their website - contacted asking why not. (they replied, but not much help-- "currently undergoing quality control processing"). Supp. figure 1A/B - Mol. Sieving NOT added as the trimer is stated to have a M.wt of 500kDa (which is about right) - yet the M. wt markers only go up go 245, and the band on SDS/PAGE is nowhere near even that value. jeremy.luban@umassmed.edu 29-Mar-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Yurkovetskiy L., Wang X., Pascal KE., Tomkins-Tinch C., Nyalile TP., Wang Y., Baum A., Diehl WE., Dauphin A., Carbone C., Veinotte K., Egri SB., Schaffner SF., Lemieux JE., Munro JB., Rafique A., Barve A., Sabeti PC., Kyratsous CA., Dudkina NV., Shen K., Luban J. Virus - Publications including interactions involving viral proteins 17/12/20:- Addgene website NOW has constructs listed -but probably not the one we need. (as of 27/10/20 Addgene do not have the plasmids used in this publication visible on their website - contacted asking why not. (they replied, but not much help-- "currently undergoing quality control processing"). Supp. figure 1A/B - Mol. Sieving NOT added as the trimer is stated to have a M.wt of 500kDa (which is about right) - yet the M. wt markers only go up go 245, and the band on SDS/PAGE is nowhere near even that value. No information can be found from stated references for "hsACE2.hFc". jeremy.luban@umassmed.edu Accepted 2020-DEC-17 AT 11:15 GMT AT 11:15 GMT by PPORRAS Cell 2020 Only protein-protein interactions imex curation imex curation Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. Cell 2020 imex curation Yurkovetskiy L., Wang X., Pascal KE., Tomkins-Tinch C., Nyalile TP., Wang Y., Baum A., Diehl WE., Dauphin A., Carbone C., Veinotte K., Egri SB., Schaffner SF., Lemieux JE., Munro JB., Rafique A., Barve A., Sabeti PC., Kyratsous CA., Dudkina NV., Shen K., Luban J. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus 17/12/20:- Addgene website NOW has constructs listed -but probably not the one we need. (as of 27/10/20 Addgene do not have the plasmids used in this publication visible on their website - contacted asking why not. (they replied, but not much help-- "currently undergoing quality control processing"). Supp. figure 1A/B - Mol. Sieving NOT added as the trimer is stated to have a M.wt of 500kDa (which is about right) - yet the M. wt markers only go up go 245, and the band on SDS/PAGE is nowhere near even that value. jeremy.luban@umassmed.edu 29-Mar-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 3d-em 3D electron microscopy 3D-EM electron tomog structure determination identification by structure determination identification by structure determination structure determination Yurkovetskiy L., Wang X., Pascal KE., Tomkins-Tinch C., Nyalile TP., Wang Y., Baum A., Diehl WE., Dauphin A., Carbone C., Veinotte K., Egri SB., Schaffner SF., Lemieux JE., Munro JB., Rafique A., Barve A., Sabeti PC., Kyratsous CA., Dudkina NV., Shen K., Luban J. Virus - Publications including interactions involving viral proteins 17/12/20:- Addgene website NOW has constructs listed -but probably not the one we need. (as of 27/10/20 Addgene do not have the plasmids used in this publication visible on their website - contacted asking why not. (they replied, but not much help-- "currently undergoing quality control processing"). Supp. figure 1A/B - Mol. Sieving NOT added as the trimer is stated to have a M.wt of 500kDa (which is about right) - yet the M. wt markers only go up go 245, and the band on SDS/PAGE is nowhere near even that value. Mutation information was adcquired from the related PDB entry 6xs6. jeremy.luban@umassmed.edu Accepted 2020-DEC-17 AT 11:15 GMT AT 11:15 GMT by PPORRAS Cell 2020 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 ace2-70 1 3 unspecified role unspecified role bait bait purified purified region fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey purified purified ecd_region sufficient to bind sufficient binding region sufficient to bind range range range range P0DTC2:p.Asp614Gly mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D G human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction 4 25 degrees C ace2-s-70 1 3 unspecified role unspecified role bait bait purified purified region fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey purified purified ecd_region sufficient to bind sufficient binding region sufficient to bind range range range range P0DTC2:p.Asp614Gly mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D G human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction 4 37 degrees C s-58 2 4 unspecified role unspecified role neutral component neutral component purified purified ecd_region sufficient to bind sufficient binding region sufficient to bind range range range range synthetic trimerization helix 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 P0DTC2:p.[Asp614Ala;Arg682_Arg685delinsAla;Lys986_Val987delinsProPro] mutation mutation certain certain sequence position certain certain certain sequence position certain KV PP certain certain sequence position certain certain certain sequence position certain RRAR A certain certain sequence position certain certain certain sequence position certain D G human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f direct interaction direct interaction Table 1 The SARS2 Spike protein, as described in this PDBe entry (6xs6), has several extra tags and mutations NOT contained in the stated plasmids used in this publication as entered on the Addgene website (plasmids #158074-7; notably C terminal His and trimerization tags and a p.[ Lys986_Val987delinsProPro;Arg682_Arg685delinsAla] mutation.) According to the Methods, the Spike proteins used are "soluble SARS-CoV-2 S Protein trimers -encoding His-tagged SARS-CoV-2 S protein", and probably contains the "ecd" = aas 1-~1213, rather than the full length or truncated (aas 1-1246) untagged constructs as described on the Addgene website. The authors have not replied to a request for clarification.