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/ Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. J. Biol. Chem. (0021-9258) 2010 imex curation Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro ion mobility mass spec ion mobility mass spectrometry of complexes ion mobility mass spec predetermined predetermined participant predetermined Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr Accepted 2013-DEC-09 by PPORRAS J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. J. Biol. Chem. (0021-9258) 2010 imex curation Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr Accepted 2013-DEC-09 by PPORRAS J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. J. Biol. Chem. (0021-9258) 2010 imex curation Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. The experiment was started under anaerobic conditions. Then the solution was exposed to light which initiated oxidation of FADH2 to FAD. After removal of unbound FAD, FAD was measured spectrophotometrically and the original FADH2 concentration was calculated based on the FAD concentration. mfontecave@cea.fr,sollagnier@cea.fr Accepted 2013-DEC-09 by PPORRAS J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. Iron-sulfur (Fe-S) cluster assembly: the SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor. J. Biol. Chem. (0021-9258) 2010 imex curation Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. mfontecave@cea.fr,sollagnier@cea.fr 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro biophysical biophysical predetermined predetermined participant predetermined Wollers S., Layer G., Garcia-Serres R., Signor L., Clemancey M., Latour J.M., Fontecave M., Ollagnier de Choudens S. The interaction detection method is Mössbauer spectroscopy. The observed transmission intensity of a gamma ray passing through a crystal drops if an interaction is present. Here Mössbauer spectroscopy measures the interaction of a Fe-S cluster with the SufB protein or SufBCD complex. mfontecave@cea.fr,sollagnier@cea.fr Accepted 2013-DEC-09 by PPORRAS J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation fadh(.) FADH(.) flavin adenine dinucleotide semiquinone radical small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) KKQJCLIXDLBMNI-UYBVJOGSSA-N KKQJCLIXDLBMNI-UYBVJOGSSA-N sufb_ecoli FeS cluster assembly protein SufB ynhE sufB b1683 JW5273 protein protein ecoli Escherichia coli (strain K12) MSRNTEATDDVKTWTGGPLNYKEGFFTQLATDELAKGINEEVVRAISAKRNEPEWMLEFRLNAYRAWLEMEEPHWLKAHYDKLNYQDYSYYSAPSCGNCDDTCASEPGAVQQTGANAFLSKEVEAAFEQLGVPVREGKEVAVDAIFDSVSVATTYREKLAEQGIIFCSFGEAIHDHPELVRKYLGTVVPGNDNFFAALNAAVASDGTFIYVPKGVRCPMELSTYFRINAEKTGQFERTILVADEDSYVSYIEGCSAPVRDSYQLHAAVVEVIIHKNAEVKYSTVQNWFPGDNNTGGILNFVTKRALCEGENSKMSWTQSETGSAITWKYPSCILRGDNSIGEFYSVALTSGHQQADTGTKMIHIGKNTKSTIISKGISAGHSQNSYRGLVKIMPTATNARNFTQCDSMLIGANCGAHTFPYVECRNNSAQLEHEATTSRIGEDQLFYCLQRGISEEDAISMIVNGFCKDVFSELPLEFAVEAQKLLAISLEHSVG DIP protein P77522 original sequence version: null 3BB49EEADB6C1278 fadh2 FADH2 1,5-dihydro-FAD DIHYDROFLAVINE-ADENINE DINUCLEOTIDE FADH2 flavin adenine dinucleotide (reduced) adenosine 5'-(3-{D-ribo-5-[7,8-dimethyl-2,4-dioxo-1,3,4,5-tetrahydrobenzo[g]pteridin-10(2H)-yl]-2,3,4-trihydroxypentyl} dihydrogen diphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) YPZRHBJKEMOYQH-UYBVJOGSSA-N YPZRHBJKEMOYQH-UYBVJOGSSA-N sufd_ecoli FeS cluster assembly protein SufD ynhC sufD b1681 JW1671 protein protein ecoli Escherichia coli (strain K12) MAGLPNSSNALQQWHHLFEAEGTKRSPQAQQHLQQLLRTGLPTRKHENWKYTPLEGLINSQFVSIAGEISPQQRDALALTLDSVRLVFVDGRYVPALSDATEGSGYEVSINDDRQGLPDAIQAEVFLHLTESLAQSVTHIAVKRGQRPAKPLLLMHITQGVAGEEVNTAHYRHHLDLAEGAEATVIEHFVSLNDARHFTGARFTINVAANAHLQHIKLAFENPLSHHFAHNDLLLAEDATAFSHSFLLGGAVLRHNTSTQLNGENSTLRINSLAMPVKNEVCDTRTWLEHNKGFCNSRQLHKTIVSDKGRAVFNGLINVAQHAIKTDGQMTNNNLLMGKLAEVDTKPQLEIYADDVKCSHGATVGRIDDEQIFYLRSRGINQQDAQQMIIYAFAAELTEALRDEGLKQQVLARIGQRLPGGAR DIP protein P77689 original sequence version: null 5744291F1E0AE825 fe4s4 iron-sulfur cluster Fe4S4 iron-sulfur cluster 4 iron, 4 sulfur cluster small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) sufc_ecoli Probable ATP-dependent transporter SufC ynhD sufC b1682 JW1672 protein protein ecoli Escherichia coli (strain K12) MLSIKDLHVSVEDKAILRGLSLDVHPGEVHAIMGPNGSGKSTLSATLAGREDYEVTGGTVEFKGKDLLALSPEDRAGEGIFMAFQYPVEIPGVSNQFFLQTALNAVRSYRGQETLDRFDFQDLMEEKIALLKMPEDLLTRSVNVGFSGGEKKRNDILQMAVLEPELCILDESDSGLDIDALKVVADGVNSLRDGKRSFIIVTHYQRILDYIKPDYVHVLYQGRIVKSGDFTLVKQLEEQGYGWLTEQQ DIP protein P77499 original sequence version: null 78FB1108E1850DF9 sufb-sufc2-sufd-1 2 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. S1a, S1d sufb-sufc1-sufd-1 2 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. S1a, S1d sufb-sufc2-sufd-2 1 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. S1b, S1c sufb-sufc1-sufd-2 1 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. S1b, S1c sufbcd-fadh2-1 3 7 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. 3b, 4 sufbcd-fadh-1 3 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. 3b sufb-sufc-sufd 4 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role neutral component neutral component 57fe-s radiolabelled radiolabel radiolabeled radiolabelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 2b, S3 The iron in the Fe-S cluster is radiolabelled 57Fe. sufb-fe_s 4 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role neutral component neutral component 57fe-s radiolabelled radiolabel radiolabeled radiolabelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 2a The iron in the Fe-S cluster is radiolabelled 57Fe.