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/ gazda-2013-6 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro crosslink cross-linking study crosslink weight by comassie molecular weight estimation by coomasie staining weight by comassie Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. Proteins containing the UV-inducible crosslinking amino acid p-benzoyl-L-phenylalanine were UV-irradiated for 15-30 minutes. Protein complexes separated by SEC. thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-2 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-4 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. caeel Caenorhabditis elegans anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-5 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro crosslink cross-linking study crosslink weight by comassie molecular weight estimation by coomasie staining weight by comassie Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. Proteins containing the UV-inducible crosslinking amino acid p-benzoyl-L-phenylalanine were UV-irradiated for 15-30 minutes thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-1 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-3 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation gazda-2013-7 The myosin chaperone UNC-45 is organized in tandem modules to support myofilament formation in C. elegans. in vitro In vitro crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Gazda L., Pokrzywa W., Hellerschmied D., Löwe T., Forné I., Mueller-Planitz F., Hoppe T., Clausen T. Proteins containing the UV-inducible crosslinking amino acid p-benzoyl-L-phenylalanine were UV-irradiated for 15-30 minutes thorsten.hoppe@uni-koeln.de, clausen@imp.univie.ac.at Cell (0092-8674) 2013 Only protein-protein interactions imex curation g5eg62_caeel unc-45 CELE_F30H5.1 F30H5.1 protein protein caeel Caenorhabditis elegans MVARVQTAEEIRDEGNAAVKDQDYIKADELYTEALQLTTDEDKALRPVLYRNRAMARLKRDDFEGAQSDCTKALEFDGADVKALFRRSLAREQLGNVGPAFQDAKEALRLSPNDKGIVEVLQRLVKANNDKIKQTTSLANKVTDMEKLAFRGEAKDTEQKMTALNNLLVLCRESESGATGVWNQGALVPFVLNLINDASENEEVTVTAIRILDETIKNSVRCMKFLAMHDPDGPKSVRFVCRLMCKKSTKDFVDATGILVQRVFNAMAKMDRQKEMKPDPEVAEANKIWIIRVLLELQEMLQDPKVGAVQRETCIDLFLKNLMHMDGGIPRGWSWKFVEERGLLALLDVASQIPELCEYPVSAETRQHVAICLQRLEEDMVFDTKRTIFKEKVDMFFNALISRCTNDDEGHKYRIKLSCFLITMLQGPVDIGINLITNDQLTPIMLEMAASQDHLMQGIAAELIVATVSKHERAINMLKVGIPVLRALYDSEDPTVKVRALVGLCKIGAAGGDDISKATMKEEAVISLAKTCKKFLLETEKYSVDIRRYACEGLSYLSLDADVKEWIVDDSLLLKALVLLAKKAGALCVYTLATIYANLSNAFEKPKVDEEMVKLAQFAKHHVPETHPKDTEEYVEKRVRALVEEGAVPACVAVSKTESKNALELIARSLLAFAEYEDLRGRIIAEGGTVLCLRLTKEASGEGKIKAGHAIAKLGAKADPMISFPGQRAYEVVKPLCDLLHPDVEGKANYDSLLTLTNLASVSDSIRGRILKEKAIPKIEEFWFMTDHEHLRAAAAELLLNLLFFEKFYEETVAPGTDRLKLWVLYSAEVEEERLSRASAAGFAILTEDENACARIMDEIKSWPEVFKDIAMHEDAETQRRGLMGIANIMHSSNKLCSEIVSSEVFRVLVAVTKLGTINQERAGSTEQAKRGLEAAEKFGLIKATDREIYERENQMSTIQE hsp90_caeel Heat shock protein 90 daf-21 Abnormal dauer formation protein 21 C47E8.5 protein protein caeel Caenorhabditis elegans MSENAETFAFQAEIAQLMSLIINTFYSNKEIYLRELISNASDALDKIRYQALTEPSELDTGKELFIKITPNKEEKTLTIMDTGIGMTKADLVNNLGTIAKSGTKAFMEALQAGADISMIGQFGVGFYSAFLVADKVVVTSKNNDDDSYQWESSAGGSFVVRPFNDPEVTRGTKIVMHIKEDQIDFLEERKIKEIVKKHSQFIGYPIKLVVEKEREKEVEDEEAVEAKDEEKKEGEVENVADDADKKKTKKIKEKYFEDEELNKTKPIWTRNPDDISNEEYAEFYKSLSNDWEDHLAVKHFSVEGQLEFRALLFVPQRAPFDLFENKKSKNSIKLYVRRVFIMENCEELMPEYLNFIKGVVDSEDLPLNISREMLQQSKILKVIRKNLVKKCMELIDEVAEDKDNFKKFYEQFGKNLKLGIHEDSTNRKKLSDFLRYSTSAGDEPTSLKEYVSRMKENQTQIYYITGESKDVVAASAFVERVKSRGFEVLYMCDPIDEYCVQQLKEYDGKKLVSVTKEGLELPETEEEKKKFEEDKVAYENLCKVIKDILEKKVEKVGVSNRLVSSPCCIVTSEYGWSANMERIMKAQALRDSSTMGYMAAKKHLEINPDHAIMKTLRDRVEVDKNDKTVKDLVVLLFETALLASGFSLEEPQSHASRIYRMIKLGLDIGDDEIEDSAVPSSCTAEAKIEGAEEDASRMEEVD hsp1_caeel Heat shock protein hsp-1 hsp-1 hsp70a F26D10.3 Heat shock 70 kDa protein A protein protein caeel Caenorhabditis elegans MSKHNAVGIDLGTTYSCVGVFMHGKVEIIANDQGNRTTPSYVAFTDTERLIGDAAKNQVAMNPHNTVFDAKRLIGRKFDDPAVQSDMKHWPFKVISAEGAKPKVQVEYKGENKIFTPEEISSMVLLKMKETAEAFLGTTVKDAVVTVPAYFNDSQRQATKDAGAIAGLNVLRIINEPTAAAIAYGLDKKGHGERNVLIFDLGGGTFDVSILTIEDGIFEVKSTAGDTHLGGEDFDNRMVNHFCAEFKRKHKKDLASNPRALRRLRTACERAKRTLSSSSQASIEIDSLFEGIDFYTNITRARFEELCADLFRSTMDPVEKSLRDAKMDKSQVHDIVLVGGSTRIPKVQKLLSDLFSGKELNKSINPDEAVAYGAAVQAAILSGDKSEAVQDLLLLDVAPLSLGIETAGGVMTALIKRNTTIPTKTAQTFTTYSDNQPGVLIQVYEGERAMTKDNNLLGKFELSGIPPAPRGVPQIEVTFDIDANGILNVSATDKSTGKQNKITITNDKGRLSKDDIERMVNEAEKYKADDEAQKDRIGAKNGLESYAFNLKQTIEDEKLKDKISPEDKKKIEDKCDEILKWLDSNQTAEKEEFEHQQKDLEGLANPIISKLYQSAGGAPPGAAPGGAAGGAGGPTIEEVD myo4_caeel Myosin-4 unc-54 myo-4 Myosin heavy chain B Uncoordinated protein 54 F11C3.3 protein protein caeel Caenorhabditis elegans MEHEKDPGWQYLRRTREQVLEDQSKPYDSKKNVWIPDPEEGYLAGEITATKGDQVTIVTARGNEVTLKKELVQEMNPPKFEKTEDMSNLSFLNDASVLHNLRSRYAAMLIYTYSGLFCVVINPYKRLPIYTDSCARMFMGKRKTEMPPHLFAVSDEAYRNMLQDHENQSMLITGESGAGKTENTKKVICYFAAVGASQQEGGAEVDPNKKKVTLEDQIVQTNPVLEAFGNAKTVRNNNSSRFGKFIRIHFNKHGRLASCDIEHYLLEKSRVIRQAPGERCYHIFYQIYSDFRPELKKELLLDLPIKDYWFVAQAELIIDGIDDVEEFQLTDEAFDILNFSAVEKQDCYRLMSAHMHMGNMKFKQRPREEQAEPDGTDEAEKASNMYGIGCEEFLKALTKPRVKVGTEWVSKGQNCEQVNWAVGAMAKGLYSRVFNWLVKKCNLTLDQKGIDRDYFIGVLDIAGFEIFDFNSFEQLWINFVNEKLQQFFNHHMFVLEQEEYAREGIQWVFIDFGLDLQACIELIEKPLGIISMLDEECIVPKATDLTLASKLVDQHLGKHPNFEKPKPPKGKQGEAHFAMRHYAGTVRYNCLNWLEKNKDPLNDTVVSAMKQSKGNDLLVEIWQDYTTQEEAAAKAKEGGGGGKKKGKSGSFMTVSMLYRESLNNLMTMLNKTHPHFIRCIIPNEKKQSGMIDAALVLNQLTCNGVLEGIRICRKGFPNRTLHPDFVQRYAILAAKEAKSDDDKKKCAEAIMSKLVNDGSLSEEMFRIGLTKVFFKAGVLAHLEDIRDEKLATILTGFQSQIRWHLGLKDRKRRMEQRAGLLIVQRNVRSWCTLRTWEWFKLYGKVKPMLKAGKEAEELEKINDKVKALEDSLAKEEKLRKELEESSAKLVEEKTSLFTNLESTKTQLSDAEERLAKLEAQQKDASKQLSELNDQLADNEDRTADVQRAKKKIEAEVEALKKQIQDLEMSLRKAESEKQSKDHQIRSLQDEMQQQDEAIAKLNKEKKHQEEINRKLMEDLQSEEDKGNHQNKVKAKLEQTLDDLEDSLEREKRARADLDKQKRKVEGELKIAQENIDESGRQRHDLENNLKKKESELHSVSSRLEDEQALVSKLQRQIKDGQSRISELEEELENERQSRSKADRAKSDLQRELEELGEKLDEQGGATAAQVEVNKKREAELAKLRRDLEEANMNHENQLGGLRKKHTDAVAELTDQLDQLNKAKAKVEKDKAQAVRDAEDLAAQLDQETSGKLNNEKLAKQFELQLTELQSKADEQSRQLQDFTSLKGRLHSENGDLVRQLEDAESQVNQLTRLKSQLTSQLEEARRTADEEARERQTVAAQAKNYQHEAEQLQESLEEEIEGKNEILRQLSKANADIQQWKARFEGEGLLKADELEDAKRRQAQKINELQEALDAANSKNASLEKTKSRLVGDLDDAQVDVERANGVASALEKKQKGFDKIIDEWRKKTDDLAAELDGAQRDLRNTSTDLFKAKNAQEELAEVVEGLRRENKSLSQEIKDLTDQLGEGGRSVHEMQKIIRRLEIEKEELQHALDEAEAALEAEESKVLRAQVEVSQIRSEIEKRIQEKEEEFENTRKNHARALESMQASLETEAKGKAELLRIKKKLEGDINELEIALDHANKANADAQKNLKRYQEQVRELQLQVEEEQRNGADTREQFFNAEKRATLLQSEKEELLVANEAAERARKQAEYEAADARDQANEANAQVSSLTSAKRKLEGEIQAIHADLDETLNEYKAAEERSKKAIADATRLAEELRQEQEHSQHVDRLRKGLEQQLKEIQVRLDEAEAAALKGGKKVIAKLEQRVRELESELDGEQRRFQDANKNLGRADRRVRELQFQVDEDKKNFERLQDLIDKLQQKLKTQKKQVEEAEELANLNLQKYKQLTHQLEDAEERADQAENSLSKMRSKSRASASVAPGLQSSASAAVIRSPSRARASDF unc_45-2 1389313 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 5B unc_45-2 1389313 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Fig. 5B unc_45-2 1389313 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) Stoichiometry: 3.0 direct interaction direct interaction false false false Fig. 5B unc-1 1389313 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) Stoichiometry: 5.0 direct interaction direct interaction false false false Fig. 5B daf_21-unc_45-1 1389314 n/a 1389321 unspecified role unspecified role neutral component neutral component region 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 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Lys82Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1389314 Fig. 2A hsp_1-unc_45-1 1389314 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Lys82Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1389322 unspecified role unspecified role neutral component neutral component region 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) direct interaction direct interaction false false false 1389314 Fig. 2A daf_21-unc_45-2 1389314 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 1389321 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1389314 Fig. 2A unc_45-daf_21 1389315 n/a 1389320 unspecified role unspecified role bait bait c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1389323 unspecified role unspecified role prey prey n/a 1389321 unspecified role unspecified role prey prey association association false false false Fig. 3D unc_45-daf_21-1 1389315 n/a 1389320 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false G5EG62:p.Asn758Tyr mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1389323 unspecified role unspecified role prey prey physical association physical association false false false Fig. 3D unc_45-daf_21-2 1389315 n/a 1389323 unspecified role unspecified role prey prey n/a 1389320 unspecified role unspecified role bait bait G5EG62:p.[Val480Arg;Val484Arg] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Supp. Fig. 6D unc_45 1389316 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). G5EG62:p.[Val480Arg;Val484Arg] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false G5EG62:p.Leu121Trp 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. 5A, Supp. Fig. 5A Chain formation depends strongly on UNC-45 concentration, with the longer chains occurring only at elevated protein concentration. unc_45-1 1389316 n/a 1389320 unspecified role unspecified role neutral component neutral component G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). c-terminal 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: 3.0 direct interaction direct interaction false false false Fig. 5A, Supp Fig. 5A Chain formation depends strongly on UNC-45 concentration, with the longer chains occurring only at elevated protein concentration. unc_45-2 1389316 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Fig. 5A, Supp. Fig. 5A Chain formation depends strongly on UNC-45 concentration, with the longer chains occurring only at elevated protein concentration. daf_21-unc_45 1389317 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 1389321 unspecified role unspecified role neutral component neutral component region 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) direct interaction direct interaction false false false Supp. Table 1 2.9A Rworking = 0.24 Rfree = 0.26 hsp_1-unc_45 1389317 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 1389322 unspecified role unspecified role neutral component neutral component region 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) direct interaction direct interaction false false false Supp. Table 1 3.6A Rworking = 0.23 Rfree = 0.28 daf_21-unc_45-3 1389318 n/a 1389321 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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) direct interaction direct interaction false false false Fig. 2B, Supp. Fig. 2 hsp_1-unc_45-2 1389318 n/a 1389322 unspecified role unspecified role neutral component neutral component c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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) direct interaction direct interaction false false false Fig. 2B, Supp. 2A unc_45-2 1389319 n/a 1389320 unspecified role unspecified role neutral component neutral component c-terminal 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 G5EG62:p.Gln452Xaa mutation mutation certain certain sequence position certain certain certain sequence position certain false Glutamine 452 was mutated to unconventional amino acid p-benzoyl-L-phenylalanine (pBpa). ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 5c UV-activated pBpa452 crosslinks to the Lys131/Ile132/Lys133 sequence