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/ Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. 1a in vitro In vitro pull down pull down edman degradation edman degradation Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. 1a Accepted 2010-JAN-10 by SKERRIEN (applied to all records created before 2005-Sep-01). Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. 1a in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. N terminal sequencing was used to determine the identity of the proteins. 1a Accepted 2010-JAN-10 by SKERRIEN (applied to all records created before 2005-Sep-01). Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Cys-328 of IscS and Cys-63 of IscU are the sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of iron-sulfur cluster assembly. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. 1a in vitro In vitro pull down pull down edman degradation edman degradation Kato S., Mihara H., Kurihara T., Takahashi Y., Tokumoto U., Yoshimura T., Esaki N. N terminal sequencing was used to determine the identity of the proteins. 1a Accepted 2010-JAN-10 by SKERRIEN (applied to all records created before 2005-Sep-01). Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 iscs_ecoli Cysteine desulfurase IscS iscS b2530 JW2514 yfhO yzzO ThiI transpersulfidase NifS protein homolog TusA transpersulfidase nuvC protein protein ecoli Escherichia coli (strain K12) MKLPIYLDYSATTPVDPRVAEKMMQFMTMDGTFGNPASRSHRFGWQAEEAVDIARNQIADLVGADPREIVFTSGATESDNLAIKGAANFYQKKGKHIITSKTEHKAVLDTCRQLEREGFEVTYLAPQRNGIIDLKELEAAMRDDTILVSIMHVNNEIGVVQDIAAIGEMCRARGIIYHVDATQSVGKLPIDLSQLKVDLMSFSGHKIYGPKGIGALYVRRKPRVRIEAQMHGGGHERGMRSGTLPVHQIVGMGEAYRIAKEEMATEMERLRGLRNRLWNGIKDIEEVYLNGDLEHGAPNILNVSFNYVEGESLIMALKDLAVSSGSACTSASLEPSYVLRALGLNDELAHSSIRFSLGRFTTEEEIDYTIELVRKSIGRLRDLSPLWEMYKQGVDLNSIEWAHH 1E4F2E6F9CD266B0 1E4F2E6F9CD266B0 iscu_ecoli Iron-sulfur cluster assembly scaffold protein IscU iscU nifU yfhN JW2513 b2529 Sulfur acceptor protein IscU protein protein ecoli Escherichia coli (strain K12) MAYSEKVIDHYENPRNVGSFDNNDENVGSGMVGAPACGDVMKLQIKVNDEGIIEDARFKTYGCGSAIASSSLVTEWVKGKSLDEAQAIKNTDIAEELELPPVKIHCSILAEDAIKAAIADYKSKREAK DIP protein P0ACD4 original sequence version: 43 DIP protein P0ACD4 original sequence version: null 47516021442B6535 iscu-iscs 1 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) covalent binding covalent binding The proteins are linked by a disulfide bond as the complex is sensitive to beta-mercaptoethanol bait and prey are overexpressed iscu-iscs-1 3 4 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 ecobb Escherichia coli (strain B / BL21) physical association physical association stoichiometry is 1:1. size of the complex was 64kDa. proteins over expressed iscu-iscu 3 5 unspecified role unspecified role prey prey 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 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 ecobb Escherichia coli (strain B / BL21) physical association physical association stoichiometry is 1:1. size of the complex was 38kDa. proteins overexpressed. iscu-iscs-3 2 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 cys 63 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey cys 328 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 22 24 physical association physical association Size of the complex was 150kDa. Interacting Cys were identified by cyanolysis, identification of the crosslinked residues was done by digestion of the complex followed by reducing the disulfide bond and detection by ABD-f labelling. Mutation analysis indicated that though the disulfide bond formed was between Cys-328 of Iscs and Cys-63 of IscU the specific interaction did not require these residues. iscu-iscu-1 2 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role prey prey 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 physical association physical association stoichiometry is 1:1. size of the complex was 45kDa. iscu-iscu-4 1 5 unspecified role unspecified role bait bait c terminus 6 his tag 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 ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) covalent binding covalent binding The proteins are linked by a disulfide bond as the complex is sensitive to beta-mercaptoethanol homodimer proteins are overexpressed