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/ vergnes-2006-1 NarJ chaperone binds on two distinct sites of the aponitrate reductase of Escherichia coli to coordinate molybdenum cofactor insertion and assembly. ecoli Escherichia coli (strain K12) 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Vergnes A., Pommier J., Toci R., Blasco F., Giordano G., Magalon A. Identity of the T12 and T25 fusion proteins were obtained from reference 24 -Magalon et al,. 2002. magalon@ibsm.cnrs-mrs.fr. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation vergnes-2006-2 NarJ chaperone binds on two distinct sites of the aponitrate reductase of Escherichia coli to coordinate molybdenum cofactor insertion and assembly. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Vergnes A., Pommier J., Toci R., Blasco F., Giordano G., Magalon A. Identity of the T12 and T25 fusion proteins were obtained from reference 24 -Magalon et al,. 2002. magalon@ibsm.cnrs-mrs.fr. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation narg_ecoli Respiratory nitrate reductase 1 alpha chain narG bisD narC b1224 Nitrate reductase A subunit alpha Quinol-nitrate oxidoreductase subunit alpha JW1215 protein protein ecoli Escherichia coli (strain K12) MSKFLDRFRYFKQKGETFADGHGQLLNTNRDWEDGYRQRWQHDKIVRSTHGVNCTGSCSWKIYVKNGLVTWETQQTDYPRTRPDLPNHEPRGCPRGASYSWYLYSANRLKYPMMRKRLMKMWREAKALHSDPVEAWASIIEDADKAKSFKQARGRGGFVRSSWQEVNELIAASNVYTIKNYGPDRVAGFSPIPAMSMVSYASGARYLSLIGGTCLSFYDWYCDLPPASPQTWGEQTDVPESADWYNSSYIIAWGSNVPQTRTPDAHFFTEVRYKGTKTVAVTPDYAEIAKLCDLWLAPKQGTDAAMALAMGHVMLREFHLDNPSQYFTDYVRRYTDMPMLVMLEERDGYYAAGRMLRAADLVDALGQENNPEWKTVAFNTNGEMVAPNGSIGFRWGEKGKWNLEQRDGKTGEETELQLSLLGSQDEIAEVGFPYFGGDGTEHFNKVELENVLLHKLPVKRLQLADGSTALVTTVYDLTLANYGLERGLNDVNCATSYDDVKAYTPAWAEQITGVSRSQIIRIAREFADNADKTHGRSMIIVGAGLNHWYHLDMNYRGLINMLIFCGCVGQSGGGWAHYVGQEKLRPQTGWQPLAFALDWQRPARHMNSTSYFYNHSSQWRYETVTAEELLSPMADKSRYTGHLIDFNVRAERMGWLPSAPQLGTNPLTIAGEAEKAGMNPVDYTVKSLKEGSIRFAAEQPENGKNHPRNLFIWRSNLLGSSGKGHEFMLKYLLGTEHGIQGKDLGQQGGVKPEEVDWQDNGLEGKLDLVVTLDFRLSSTCLYSDIILPTATWYEKDDMNTSDMHPFIHPLSAAVDPAWEAKSDWEIYKAIAKKFSEVCVGHLGKETDIVTLPIQHDSAAELAQPLDVKDWKKGECDLIPGKTAPHIMVVERDYPATYERFTSIGPLMEKIGNGGKGIAWNTQSEMDLLRKLNYTKAEGPAKGQPMLNTAIDAAEMILTLAPETNGQVAVKAWAALSEFTGRDHTHLALNKEDEKIRFRDIQAQPRKIISSPTWSGLEDEHVSYNAGYTNVHELIPWRTLSGRQQLYQDHQWMRDFGESLLVYRPPIDTRSVKEVIGQKSNGNQEKALNFLTPHQKWGIHSTYSDNLLMLTLGRGGPVVWLSEADAKDLGIADNDWIEVFNSNGALTARAVVSQRVPAGMTMMYHAQERIVNLPGSEITQQRGGIHNSVTRITPKPTHMIGGYAHLAYGFNYYGTVGSNRDEFVVVRKMKNIDWLDGEGNDQVQESVK narj_ecoli Nitrate reductase molybdenum cofactor assembly chaperone NarJ narJ b1226 JW1217 Redox enzyme maturation protein NarJ protein protein ecoli Escherichia coli (strain K12) MIELVIVSRLLEYPDAALWQHQQEMFEAIAASKNLPKEDAHALGIFLRDLTTMDPLDAQAQYSELFDRGRATSLLLFEHVHGESRDRGQAMVDLLAQYEQHGLQLNSRELPDHLPLYLEYLAQLPQSEAVEGLKDIAPILALLSARLQQRESRYAVLFDLLLKLANTAIDSDKVAEKIADEARDDTPQALDAVWEEEQVKFFADKGCGDSAITAHQRRFAGAVAPQYLNITTGGQH 17306C34A57DF1E2 narh_ecoli Respiratory nitrate reductase 1 beta chain narH b1225 Nitrate reductase A subunit beta Quinol-nitrate oxidoreductase subunit beta JW1216 protein protein ecoli Escherichia coli (strain K12) MKIRSQVGMVLNLDKCIGCHTCSVTCKNVWTSREGVEYAWFNNVETKPGQGFPTDWENQEKYKGGWIRKINGKLQPRMGNRAMLLGKIFANPHLPGIDDYYEPFDFDYQNLHTAPEGSKSQPIARPRSLITGERMAKIEKGPNWEDDLGGEFDKLAKDKNFDNIQKAMYSQFENTFMMYLPRLCEHCLNPACVATCPSGAIYKREEDGIVLIDQDKCRGWRMCITGCPYKKIYFNWKSGKSEKCIFCYPRIEAGQPTVCSETCVGRIRYLGVLLYDADAIERAASTENEKDLYQRQLDVFLDPNDPKVIEQAIKDGIPLSVIEAAQQSPVYKMAMEWKLALPLHPEYRTLPMVWYVPPLSPIQSAADAGELGSNGILPDVESLRIPVQYLANLLTAGDTKPVLRALKRMLAMRHYKRAETVDGKVDTRALEEVGLTEAQAQEMYRYLAIANYEDRFVVPSSHRELAREAFPEKNGCGFTFGDGCHGSDTKFNLFNSRRIDAIDVTSKTEPHP narg-narj-7 1143452 n/a 1143454 unspecified role unspecified role bait bait cyaa_t18_domain tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1143455 unspecified role unspecified role prey prey physical association physical association false false false Fig 4A Unexpectedly, deletion of the first 40 residues of NarG does not hamper its interaction with NarJ. narg-narh-8 1143452 n/a 1143454 unspecified role unspecified role bait bait cyaa_t18_domain tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1143456 unspecified role unspecified role prey prey physical association physical association false false false Fig 4A NarG (aa 1-40) binds NarH, but very weakly compared to the NArg(1-40) NarJ reaction. Similarly the NarG construct (aa 41-1247) also binds to NarH, but by a slightly reduced amount. This interaction gave a result of 45 Miller units as compared to the negative control that gave a result of 50 Miller units. narj-narg-5 1143453 n/a 1143455 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1143454 unspecified role unspecified role prey prey cyaa_t18_domain tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 4B narj-narg-6 1143453 n/a 1143454 unspecified role unspecified role prey prey cyaa_t25_domain tag 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 n/a 1143455 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 4C