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/ morimoto-2003-1 A HEAT-repeats containing protein, IaiH, stabilizes the iron-sulfur cluster bound to the cyanobacterial IscA homologue, IscA2. ecoli Escherichia coli (strain K12) anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Morimoto K., Sato S., Tabata S., Nakai M. Cyanobacteria - Interaction dataset based on Cyanobacteria proteins and related species fig 4 nakai@protein.osaka-u.ac.jp J. Biochem. (0021-924X) 2003 morimoto-2003-2 A HEAT-repeats containing protein, IaiH, stabilizes the iron-sulfur cluster bound to the cyanobacterial IscA homologue, IscA2. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 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 Morimoto K., Sato S., Tabata S., Nakai M. Cyanobacteria - Interaction dataset based on Cyanobacteria proteins and related species fig 4 nakai@protein.osaka-u.ac.jp J. Biochem. (0021-924X) 2003 morimoto-2003-3 A HEAT-repeats containing protein, IaiH, stabilizes the iron-sulfur cluster bound to the cyanobacterial IscA homologue, IscA2. in vitro In vitro biophysical biophysical predetermined predetermined participant predetermined Morimoto K., Sato S., Tabata S., Nakai M. Cyanobacteria - Interaction dataset based on Cyanobacteria proteins and related species Spectroscopic properties of the complex while adding EDTA or DTT (250nm-550 nm). fig 4 nakai@protein.osaka-u.ac.jp J. Biochem. (0021-924X) 2003 p72742_syny3 slr1098 protein protein syny3 Synechocystis sp. (strain PCC 6803 / Kazusa) MRKYGAMRNQDDLLLEDSWVDPLDQLDDDVAPPPPDPDEMLVLLTSREAPQRMLAARAFCEIADRRAVEPLINLLGDSCPLVRVSAAYALGRNTDPTIVEPLIQSLQTDFNGYVRKGLVWALGNCGDRRALAPLIHALKTDISAVRLWSASSLPNIAKLAYEDVIATIPPLIEGLRRDKMAAVRSNCAWAVGQLCRELPSNVVYATAIDALLEALVEDEDFGVKEDAKSALLRLGDARGLQMIEDLEMDGLI y1565_syny3 Uncharacterized protein slr1565 slr1565 protein protein syny3 Synechocystis sp. (strain PCC 6803 / Kazusa) MLQLTPSAAQEIKRLQHSRQLTRHHFRLAVRPGGCAGWLYHLDFVPEITADDLEYESGGVTVLVDSQSAGYLHNLKLDYAEDLMGGGFRFTNPNAAQVCSCSLSFAPNLEKNL iaih-isca2-1 Co-IP: bait = IaiH, prey = IscA2 257299 N/A 257302 unspecified role unspecified role bait bait N/A 257303 unspecified role unspecified role prey prey P74596:p.Cys35Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P74596:p.Cys99Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false iaih-isca2-2 Y2H: bait =IscA2, prey = IaiH 257300 N/A 257303 unspecified role unspecified role bait bait P74596:p.Cys99Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P74596:p.Cys35Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P74596:p.Cys101Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false N/A 257302 unspecified role unspecified role prey prey heat repeat h1 h2 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The fragments of IaiH binding IscA2 all contain two HEAT repeats designated H1 and H2 which seem to be required. iaih-isca2-5 Spectroscopic studies of IscA2/ IaiH complex 257301 N/A 257302 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) N/A 257303 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association false false false The absorption spectrum of the Isca2/IaiH complex was unaffected by the addition of EDTA. By comparison, using Isca2 alone, the intensity of the visible absorption spectrum characteristic of the [2Fe-2S] cluster decreased slightly. [2Fe-2S] cluster is stable in presence of a metal chelator but this stability is greater in presence of IaiH. The absorption spectrum of the [2Fe-2S] cluster containing Isca2/IaiH complex was unaffected by DTT or by subsequent air oxidation. By comparision, the [2Fe-2S] cluster is disassociated from the protein upon reduction by DTT. The following mutants of Isca2 C35S, C99S, C101S in complex with IaiH are reductively labil. The following mutations of Isca2 C35S, C101S both affect the absorption sprectrum of the Isca2/IaiH complex at 420-460 nm.