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/ muramatsu-2016-3 in vitro In vitro protease assay protease assay weight by comassie molecular weight estimation by coomasie staining weight by comassie Muramatsu T., Takemoto C., Kim YT., Wang H., Nishii W., Terada T., Shirouzu M., Yokoyama S. Virus - Publications including interactions involving viral proteins tomonari.muramatsu@riken.jp,yokoyama@riken.jp Proceedings of the National Academy of Sciences of the United States of America 2016 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus muramatsu-2016-2 in vitro In vitro protease assay protease assay tag fluorescence tag visualisation by fluorescence tag fluorescence Muramatsu T., Takemoto C., Kim YT., Wang H., Nishii W., Terada T., Shirouzu M., Yokoyama S. Virus - Publications including interactions involving viral proteins The substrate is labelled with N-methyl-anthranilic acid (NMA), a fluorescent probe for peptide commonly used in the study of proteases. Conjugation of NMA via its amino group to a peptide substrate results in quenching of the fluorescent signal. Upon enzymatic cleavage of the peptide by proteases, NMA is released and its fluorescence can be used to quantify enzyme activity. tomonari.muramatsu@riken.jp,yokoyama@riken.jp Proceedings of the National Academy of Sciences of the United States of America 2016 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus muramatsu-2016-1 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Muramatsu T., Takemoto C., Kim YT., Wang H., Nishii W., Terada T., Shirouzu M., Yokoyama S. Virus - Publications including interactions involving viral proteins The protein had been synthesised in E. coli cell-free protein synthesis system tomonari.muramatsu@riken.jp,yokoyama@riken.jp Proceedings of the National Academy of Sciences of the United States of America 2016 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus r1a_sars Replicase polyprotein 1a ORF1a polyprotein 1a protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus MESLVLGVNEKTHVQLSLPVLQVRDVLVRGFGDSVEEALSEAREHLKNGTCGLVELEKGVLPQLEQPYVFIKRSDALSTNHGHKVVELVAEMDGIQYGRSGITLGVLVPHVGETPIAYRNVLLRKNGNKGAGGHSYGIDLKSYDLGDELGTDPIEDYEQNWNTKHGSGALRELTRELNGGAVTRYVDNNFCGPDGYPLDCIKDFLARAGKSMCTLSEQLDYIESKRGVYCCRDHEHEIAWFTERSDKSYEHQTPFEIKSAKKFDTFKGECPKFVFPLNSKVKVIQPRVEKKKTEGFMGRIRSVYPVASPQECNNMHLSTLMKCNHCDEVSWQTCDFLKATCEHCGTENLVIEGPTTCGYLPTNAVVKMPCPACQDPEIGPEHSVADYHNHSNIETRLRKGGRTRCFGGCVFAYVGCYNKRAYWVPRASADIGSGHTGITGDNVETLNEDLLEILSRERVNINIVGDFHLNEEVAIILASFSASTSAFIDTIKSLDYKSFKTIVESCGNYKVTKGKPVKGAWNIGQQRSVLTPLCGFPSQAAGVIRSIFARTLDAANHSIPDLQRAAVTILDGISEQSLRLVDAMVYTSDLLTNSVIIMAYVTGGLVQQTSQWLSNLLGTTVEKLRPIFEWIEAKLSAGVEFLKDAWEILKFLITGVFDIVKGQIQVASDNIKDCVKCFIDVVNKALEMCIDQVTIAGAKLRSLNLGEVFIAQSKGLYRQCIRGKEQLQLLMPLKAPKEVTFLEGDSHDTVLTSEEVVLKNGELEALETPVDSFTNGAIVGTPVCVNGLMLLEIKDKEQYCALSPGLLATNNVFRLKGGAPIKGVTFGEDTVWEVQGYKNVRITFELDERVDKVLNEKCSVYTVESGTEVTEFACVVAEAVVKTLQPVSDLLTNMGIDLDEWSVATFYLFDDAGEENFSSRMYCSFYPPDEEEEDDAECEEEEIDETCEHEYGTEDDYQGLPLEFGASAETVRVEEEEEEDWLDDTTEQSEIEPEPEPTPEEPVNQFTGYLKLTDNVAIKCVDIVKEAQSANPMVIVNAANIHLKHGGGVAGALNKATNGAMQKESDDYIKLNGPLTVGGSCLLSGHNLAKKCLHVVGPNLNAGEDIQLLKAAYENFNSQDILLAPLLSAGIFGAKPLQSLQVCVQTVRTQVYIAVNDKALYEQVVMDYLDNLKPRVEAPKQEEPPNTEDSKTEEKSVVQKPVDVKPKIKACIDEVTTTLEETKFLTNKLLLFADINGKLYHDSQNMLRGEDMSFLEKDAPYMVGDVITSGDITCVVIPSKKAGGTTEMLSRALKKVPVDEYITTYPGQGCAGYTLEEAKTALKKCKSAFYVLPSEAPNAKEEILGTVSWNLREMLAHAEETRKLMPICMDVRAIMATIQRKYKGIKIQEGIVDYGVRFFFYTSKEPVASIITKLNSLNEPLVTMPIGYVTHGFNLEEAARCMRSLKAPAVVSVSSPDAVTTYNGYLTSSSKTSEEHFVETVSLAGSYRDWSYSGQRTELGVEFLKRGDKIVYHTLESPVEFHLDGEVLSLDKLKSLLSLREVKTIKVFTTVDNTNLHTQLVDMSMTYGQQFGPTYLDGADVTKIKPHVNHEGKTFFVLPSDDTLRSEAFEYYHTLDESFLGRYMSALNHTKKWKFPQVGGLTSIKWADNNCYLSSVLLALQQLEVKFNAPALQEAYYRARAGDAANFCALILAYSNKTVGELGDVRETMTHLLQHANLESAKRVLNVVCKHCGQKTTTLTGVEAVMYMGTLSYDNLKTGVSIPCVCGRDATQYLVQQESSFVMMSAPPAEYKLQQGTFLCANEYTGNYQCGHYTHITAKETLYRIDGAHLTKMSEYKGPVTDVFYKETSYTTTIKPVSYKLDGVTYTEIEPKLDGYYKKDNAYYTEQPIDLVPTQPLPNASFDNFKLTCSNTKFADDLNQMTGFTKPASRELSVTFFPDLNGDVVAIDYRHYSASFKKGAKLLHKPIVWHINQATTKTTFKPNTWCLRCLWSTKPVDTSNSFEVLAVEDTQGMDNLACESQQPTSEEVVENPTIQKEVIECDVKTTEVVGNVILKPSDEGVKVTQELGHEDLMAAYVENTSITIKKPNELSLALGLKTIATHGIAAINSVPWSKILAYVKPFLGQAAITTSNCAKRLAQRVFNNYMPYVFTLLFQLCTFTKSTNSRIRASLPTTIAKNSVKSVAKLCLDAGINYVKSPKFSKLFTIAMWLLLLSICLGSLICVTAAFGVLLSNFGAPSYCNGVRELYLNSSNVTTMDFCEGSFPCSICLSGLDSLDSYPALETIQVTISSYKLDLTILGLAAEWVLAYMLFTKFFYLLGLSAIMQVFFGYFASHFISNSWLMWFIISIVQMAPVSAMVRMYIFFASFYYIWKSYVHIMDGCTSSTCMMCYKRNRATRVECTTIVNGMKRSFYVYANGGRGFCKTHNWNCLNCDTFCTGSTFISDEVARDLSLQFKRPINPTDQSSYIVDSVAVKNGALHLYFDKAGQKTYERHPLSHFVNLDNLRANNTKGSLPINVIVFDGKSKCDESASKSASVYYSQLMCQPILLLDQALVSDVGDSTEVSVKMFDAYVDTFSATFSVPMEKLKALVATAHSELAKGVALDGVLSTFVSAARQGVVDTDVDTKDVIECLKLSHHSDLEVTGDSCNNFMLTYNKVENMTPRDLGACIDCNARHINAQVAKSHNVSLIWNVKDYMSLSEQLRKQIRSAAKKNNIPFRLTCATTRQVVNVITTKISLKGGKIVSTCFKLMLKATLLCVLAALVCYIVMPVHTLSIHDGYTNEIIGYKAIQDGVTRDIISTDDCFANKHAGFDAWFSQRGGSYKNDKSCPVVAAIITREIGFIVPGLPGTVLRAINGDFLHFLPRVFSAVGNICYTPSKLIEYSDFATSACVLAAECTIFKDAMGKPVPYCYDTNLLEGSISYSELRPDTRYVLMDGSIIQFPNTYLEGSVRVVTTFDAEYCRHGTCERSEVGICLSTSGRWVLNNEHYRALSGVFCGVDAMNLIANIFTPLVQPVGALDVSASVVAGGIIAILVTCAAYYFMKFRRVFGEYNHVVAANALLFLMSFTILCLVPAYSFLPGVYSVFYLYLTFYFTNDVSFLAHLQWFAMFSPIVPFWITAIYVFCISLKHCHWFFNNYLRKRVMFNGVTFSTFEEAALCTFLLNKEMYLKLRSETLLPLTQYNRYLALYNKYKYFSGALDTTSYREAACCHLAKALNDFSNSGADVLYQPPQTSITSAVLQSGFRKMAFPSGKVEGCMVQVTCGTTTLNGLWLDDTVYCPRHVICTAEDMLNPNYEDLLIRKSNHSFLVQAGNVQLRVIGHSMQNCLLRLKVDTSNPKTPKYKFVRIQPGQTFSVLACYNGSPSGVYQCAMRPNHTIKGSFLNGSCGSVGFNIDYDCVSFCYMHHMELPTGVHAGTDLEGKFYGPFVDRQTAQAAGTDTTITLNVLAWLYAAVINGDRWFLNRFTTTLNDFNLVAMKYNYEPLTQDHVDILGPLSAQTGIAVLDMCAALKELLQNGMNGRTILGSTILEDEFTPFDVVRQCSGVTFQGKFKKIVKGTHHWMLLTFLTSLLILVQSTQWSLFFFVYENAFLPFTLGIMAIAACAMLLVKHKHAFLCLFLLPSLATVAYFNMVYMPASWVMRIMTWLELADTSLSGYRLKDCVMYASALVLLILMTARTVYDDAARRVWTLMNVITLVYKVYYGNALDQAISMWALVISVTSNYSGVVTTIMFLARAIVFVCVEYYPLLFITGNTLQCIMLVYCFLGYCCCCYFGLFCLLNRYFRLTLGVYDYLVSTQEFRYMNSQGLLPPKSSIDAFKLNIKLLGIGGKPCIKVATVQSKMSDVKCTSVVLLSVLQQLRVESSSKLWAQCVQLHNDILLAKDTTEAFEKMVSLLSVLLSMQGAVDINRLCEEMLDNRATLQAIASEFSSLPSYAAYATAQEAYEQAVANGDSEVVLKKLKKSLNVAKSEFDRDAAMQRKLEKMADQAMTQMYKQARSEDKRAKVTSAMQTMLFTMLRKLDNDALNNIINNARDGCVPLNIIPLTTAAKLMVVVPDYGTYKNTCDGNTFTYASALWEIQQVVDADSKIVQLSEINMDNSPNLAWPLIVTALRANSAVKLQNNELSPVALRQMSCAAGTTQTACTDDNALAYYNNSKGGRFVLALLSDHQDLKWARFPKSDGTGTIYTELEPPCRFVTDTPKGPKVKYLYFIKGLNNLNRGMVLGSLAATVRLQAGNATEVPANSTVLSFCAFAVDPAKAYKDYLASGGQPITNCVKMLCTHTGTGQAITVTPEANMDQESFGGASCCLYCRCHIDHPNPKGFCDLKGKYVQIPTTCANDPVGFTLRNTVCTVCGMWKGYGCSCDQLREPLMQSADASTFLNGFAV p0c6u8-pro_0000338259 3C-like proteinase nsp5 ORF1a polyprotein 1a protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus SGFRKMAFPSGKVEGCMVQVTCGTTTLNGLWLDDTVYCPRHVICTAEDMLNPNYEDLLIRKSNHSFLVQAGNVQLRVIGHSMQNCLLRLKVDTSNPKTPKYKFVRIQPGQTFSVLACYNGSPSGVYQCAMRPNHTIKGSFLNGSCGSVGFNIDYDCVSFCYMHHMELPTGVHAGTDLEGKFYGPFVDRQTAQAAGTDTTITLNVLAWLYAAVINGDRWFLNRFTTTLNDFNLVAMKYNYEPLTQDHVDILGPLSAQTGIAVLDMCAALKELLQNGMNGRTILGSTILEDEFTPFDVVRQCSGVTFQ 3241 3546 nma-tsavlqsgfrk-dnp-nh2 peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) TSAVLQSGFRK vtfq peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) VTFQ vtlq peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) VTLQ vtaq peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) VTAQ r1a_cvhsa-r1a_cvhsa-1 3709974 N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3709977 enzyme enzyme neutral component neutral component purified purified 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Gln3546Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-3 r1a_cvhsa-r1a_cvhsa-2 3709974 N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3709977 enzyme enzyme neutral component neutral component purified purified 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P0C6U8:p.Gln3240Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-b, S4-2 p0c6u8_pro_0000338259-r1a_cvhsa-4 3709974 N/A 3709978 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-1 r1a_cvhsa-r1a_cvhsa-4 3709974 N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3709977 enzyme enzyme neutral component neutral component purified purified n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 P0C6U8:p.[Gln3240Asn;Gln3546Asn] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-4 p0c6u8_pro_0000338259-r1a_cvhsa-1 3709974 N/A 3709978 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Phe3545Leu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Ala] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Asn] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-5 p0c6u8_pro_0000338259-r1a_cvhsa-2 3709974 N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Phe3545Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Ala] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Asn] 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 gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3709978 enzyme enzyme neutral component neutral component purified purified P0C6U8-PRO_0000338259:p.Phe305Leu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-6 r1a_cvhsa-r1a_cvhsa-3 3709974 N/A 3709977 enzyme enzyme neutral component neutral component purified purified 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Gln3546Asn mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Gln3546Asn] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Phe3545Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Ala] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.[Phe3545Leu;Phe3549Asn] 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 gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 P0C6U8:p.Phe3549Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3549Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Phe3243Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Figs. 2a-e, S4-7, S4-8 r1a_cvhsa-r1a_cvhsa-5 3709974 N/A 3709977 enzyme target enzyme target substrate substrate neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 ecoli Escherichia coli (strain K12) N/A 3709977 enzyme enzyme neutral component neutral component purified purified n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 P0C6U8:p.Cys3385Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false Fig. S4-0 Cys3385Ala (C145A) is a cleavage-defective active-site mutation nma_tsavlqsgfrk_dnp_nh2-r1a_cvhsa-1 3709975 N/A 3709979 enzyme target enzyme target substrate substrate neutral component neutral component purified purified n-terminus fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709977 enzyme enzyme neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Cys3385Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 protein cleavage protein cleavage false false false 3709975 3709975 Table 2 nma_tsavlqsgfrk_dnp_nh2-r1a_cvhsa-2 3709975 N/A 3709979 enzyme target enzyme target substrate substrate neutral component neutral component purified purified n-terminus fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709977 enzyme enzyme neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Cys3385Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 3709980 inhibitor inhibitor neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) protein cleavage protein cleavage false false false 3709975 3709975 3709975 Table 2 nma_tsavlqsgfrk_dnp_nh2-r1a_cvhsa-3 3709975 N/A 3709981 inhibitor inhibitor neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709979 enzyme target enzyme target substrate substrate neutral component neutral component purified purified n-terminus fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709977 enzyme enzyme neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Cys3385Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 protein cleavage protein cleavage false false false 3709975 3709975 3709975 Table 2 nma_tsavlqsgfrk_dnp_nh2-r1a_cvhsa-4 3709975 N/A 3709982 inhibitor inhibitor neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709979 enzyme target enzyme target substrate substrate neutral component neutral component purified purified n-terminus fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3709977 enzyme enzyme neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified P0C6U8:p.Cys3385Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 protein cleavage protein cleavage false false false 3709975 3709975 3709975 Table 2 r1a_cvhsa-1 3709976 N/A 3709977 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated purified purified 3CLpro proform sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P0C6U8:p.Cys3385Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 1a-c, S1a, Table S1 2.2Å Rwork: 0.206 Rfree: 0.254 r1a_cvhsa-2 3709976 N/A 3709978 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Table S1, Figures 1, S1a 1.7Å Rwork: 0.194 Rfree: 0.221