MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. EMBO J. (0261-4189) 2006 imex curation Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. 2009-01-08 Only protein-protein interactions in vitro In vitro bn-page blue native page bn-page nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. EMBO J. (0261-4189) 2006 imex curation Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom predetermined predetermined participant predetermined Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. EMBO J. (0261-4189) 2006 imex curation Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. 2009-01-08 Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. Essential role of Isd11 in mitochondrial iron-sulfur cluster synthesis on Isu scaffold proteins. EMBO J. (0261-4189) 2006 imex curation Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast tap tandem affinity purification TAP TAP tap predetermined predetermined participant predetermined Wiedemann N., Urzica E., Guiard B., Müller H., Lohaus C., Meyer HE., Ryan MT., Meisinger C., Mühlenhoff U., Lill R., Pfanner N. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation nfs1_yeast Cysteine desulfurase, mitochondrial NFS1 SPL1 YCL017C tRNA-splicing protein SPL1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLKSTATRSITRLSQVYNVPAATYRACLVSRRFYSPPAAGVKLDDNFSLETHTDIQAAAKAQASARASASGTTPDAVVASGSTAMSHAYQENTGFGTRPIYLDMQATTPTDPRVLDTMLKFYTGLYGNPHSNTHSYGWETNTAVENARAHVAKMINADPKEIIFTSGATESNNMVLKGVPRFYKKTKKHIITTRTEHKCVLEAARAMMKEGFEVTFLNVDDQGLIDLKELEDAIRPDTCLVSVMAVNNEIGVIQPIKEIGAICRKNKIYFHTDAAQAYGKIHIDVNEMNIDLLSISSHKIYGPKGIGAIYVRRRPRVRLEPLLSGGGQERGLRSGTLAPPLVAGFGEAARLMKKEFDNDQAHIKRLSDKLVKGLLSAEHTTLNGSPDHRYPGCVNVSFAYVEGESLLMALRDIALSSGSACTSASLEPSYVLHALGKDDALAHSSIRFGIGRFSTEEEVDYVVKAVSDRVKFLRELSPLWEMVQEGIDLNSIKWSGH D709E326B3921BCC D709E326B3921BCC isd11_yeast Protein ISD11 Iron-sulfur protein biogenesis, desulfurase-interacting protein 11 YER048W-A ISD11 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPGFTAPTRRQVLSLYKEFIKNANQFNNYNFREYFLSKTRTTFRKNMNQQDPKVLMNLFKEAKNDLGVLKRQSVISQMYTFDRLVVEPLQGRKH 5FC241A528BCCB2A isu1_yeast Iron sulfur cluster assembly protein 1, mitochondrial YPL135W Iron sulfur cluster scaffold protein 1 ISU1 NUA1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLPVITRFARPALMAIRPVNAMGVLRASSITKRLYHPKVIEHYTHPRNVGSLDKKLPNVGTGLVGAPACGDVMRLQIKVNDSTGVIEDVKFKTFGCGSAIASSSYMTELVQGMTLDDAAKIKNTEIAKELSLPPVKLHCSMLAEDAIKAAIKDYKSKRNTPTMLS E240845A27D2E9EE E240845A27D2E9EE isd11-nfs1-2 4 5 ms participant Identification by mass spectrometry ms participant predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tap tagged tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F6A EMBO-Journal We used a yeast strain expressing Isd11 with a tandem affinity purification (TAP) tag and performed an IgG affinity purification after lysis of mitochondria under native conditions. We detected a prominent protein band of about 50 kDa and several smaller bands in the eluate that were not found with control mitochondria lacking a TAP tag. A mass spectrometric analysis of these bands led to the identification of numerous peptides derived from Nfs1 and Isd11 (Figure 6A). homomint mint nfs1-isd11 2 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association EMBO-Journal F6G We purified Nfs1ProtA from mitochondria lysed under non-denaturing conditions by IgG affinity purification and performed a Western blot with antibodies against Isd11 and Isu1. As expected from the stable association in BN-PAGE, Isd11 was efficiently copurified with Nfs1 (Figure 6G). homomint mint nfs1-isu1 2 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association EMBO-Journal F6G We purified Nfs1ProtA from mitochondria lysed under non-denaturing conditions by IgG affinity purification and performed a Western blot with antibodies against Isd11 and Isu1. As expected from the stable association in BN-PAGE, Isd11 was efficiently copurified with Nfs1 (Figure 6G). homomint mint nfs1-isd11-1 3 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F6B EMBO-Journal To obtain evidence for an interaction of Isd11 and Nfs1 in intact mitochondria, radiolabeled Isd11 was imported into mitochondria containing Nfs1 with a protein A tag (Figure 6B). After crosslinking with m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), Nfs1ProtA was purified by IgG affinity purification. Upon elution with SDS, a radiolabeled band of about 75 kDa was released (Figure 6B, lane 2). The size of 75 kDa corresponds to a crosslinking product between Nfs1ProtA (65 kDa) and Isd11. homomint mint isd11-nfs1-3 1 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F6E F6D EMBO-Journal To determine if the ISD complex contained both Isd11 and Nsf1, radiolabeled Isd11 was imported into Nfs1ProtA mitochondria. The ISD complex was indeed shifted to a higher molecular mass compared to the corresponding wild-type complex (Figure 6D, lanes 3 and 4). The ISD complex was indeed shifted to a higher molecular mass compared to the corresponding wild-type complex (Figure 6D, lanes 3 and 4). homomint mint