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/ wada-2009-4 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation wada-2009-6 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro comigration in sds comigration in sds page comigration in sds weight by staining molecular weight estimation by staining weight by staining Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation wada-2009-1 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation wada-2009-2 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation wada-2009-3 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by staining molecular weight estimation by staining weight by staining Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation wada-2009-5 Molecular dynamism of Fe-S cluster biosynthesis implicated by the structure of the SufC(2)-SufD(2) complex. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Wada K., Sumi N., Nagai R., Iwasaki K., Sato T., Suzuki K., Hasegawa Y., Kitaoka S., Minami Y., Outten F.W., Takahashi Y., Fukuyama K. ytaka@molbiol.saitama-u.ac.jp,fukuyama@bio.sci.osaka-u.ac.jp J. Mol. Biol. (0022-2836) 2009 Only protein-protein interactions imex curation sufd_ecoli FeS cluster assembly protein SufD ynhC sufD b1681 JW1671 protein protein ecoli Escherichia coli (strain K12) MAGLPNSSNALQQWHHLFEAEGTKRSPQAQQHLQQLLRTGLPTRKHENWKYTPLEGLINSQFVSIAGEISPQQRDALALTLDSVRLVFVDGRYVPALSDATEGSGYEVSINDDRQGLPDAIQAEVFLHLTESLAQSVTHIAVKRGQRPAKPLLLMHITQGVAGEEVNTAHYRHHLDLAEGAEATVIEHFVSLNDARHFTGARFTINVAANAHLQHIKLAFENPLSHHFAHNDLLLAEDATAFSHSFLLGGAVLRHNTSTQLNGENSTLRINSLAMPVKNEVCDTRTWLEHNKGFCNSRQLHKTIVSDKGRAVFNGLINVAQHAIKTDGQMTNNNLLMGKLAEVDTKPQLEIYADDVKCSHGATVGRIDDEQIFYLRSRGINQQDAQQMIIYAFAAELTEALRDEGLKQQVLARIGQRLPGGAR sufc_ecoli Probable ATP-dependent transporter SufC ynhD sufC b1682 JW1672 protein protein ecoli Escherichia coli (strain K12) MLSIKDLHVSVEDKAILRGLSLDVHPGEVHAIMGPNGSGKSTLSATLAGREDYEVTGGTVEFKGKDLLALSPEDRAGEGIFMAFQYPVEIPGVSNQFFLQTALNAVRSYRGQETLDRFDFQDLMEEKIALLKMPEDLLTRSVNVGFSGGEKKRNDILQMAVLEPELCILDESDSGLDIDALKVVADGVNSLRDGKRSFIIVTHYQRILDYIKPDYVHVLYQGRIVKSGDFTLVKQLEEQGYGWLTEQQ sufb_ecoli FeS cluster assembly protein SufB ynhE sufB b1683 JW5273 protein protein ecoli Escherichia coli (strain K12) MSRNTEATDDVKTWTGGPLNYKEGFFTQLATDELAKGINEEVVRAISAKRNEPEWMLEFRLNAYRAWLEMEEPHWLKAHYDKLNYQDYSYYSAPSCGNCDDTCASEPGAVQQTGANAFLSKEVEAAFEQLGVPVREGKEVAVDAIFDSVSVATTYREKLAEQGIIFCSFGEAIHDHPELVRKYLGTVVPGNDNFFAALNAAVASDGTFIYVPKGVRCPMELSTYFRINAEKTGQFERTILVADEDSYVSYIEGCSAPVRDSYQLHAAVVEVIIHKNAEVKYSTVQNWFPGDNNTGGILNFVTKRALCEGENSKMSWTQSETGSAITWKYPSCILRGDNSIGEFYSVALTSGHQQADTGTKMIHIGKNTKSTIISKGISAGHSQNSYRGLVKIMPTATNARNFTQCDSMLIGANCGAHTFPYVECRNNSAQLEHEATTSRIGEDQLFYCLQRGISEEDAISMIVNGFCKDVFSELPLEFAVEAQKLLAISLEHSVG sufd-1 1445426 n/a 1445432 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) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 4c sufc-sufd-4 1445426 n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1445433 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) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 5a sufbcd-1 1445426 n/a 1445434 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1445433 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false Fig. 5a sufbcd-2 1445427 n/a 1445433 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1445434 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false Fig. 5b sufd-sufc-2 1445427 n/a 1445433 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) n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 5b sufc-sufd-1 1445428 n/a 1445433 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) Stoichiometry: 2.0 n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig.s 1a, 1b, 2, 3c, 4a R = 0.23; Rwork = 0.23; Rfree = 0.26 2.2 A sufc-sufd-2 1445429 n/a 1445433 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) Stoichiometry: 2.0 n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 1c sufc-sufd-3 1445430 n/a 1445433 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 P77499:p.Tyr86Cys mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1445432 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 3d Tyr86 was mutated to Cyc to enhance SufC homodimer interface binding in the presence of ATP and metal ions (Mg2+, Cu2+). It produced an additional, larger band on the native gel than the WT complex suggesting a conformational change in the complex quaternary structure. sufd-sufbc 1445431 n/a 1445434 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1445433 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1445432 unspecified role unspecified role bait bait 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) physical association physical association false false false Fig. 4d