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/ van-2005-2 Pam17 is required for architecture and translocation activity of the mitochondrial protein import motor. yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom western blot western blot Immuno blot Immuno blot van der Laan M., Chacinska A., Lind M., Perschil I., Sickmann A., Meyer H.E., Guiard B., Meisinger C., Pfanner N., Rehling P. Nikolaus.Pfanner@biochemie.uni-freiburg.de, Peter.Rehling@biochemie.uni-freiburg.de Mol. Cell. Biol. (0270-7306) 2005 Only protein-protein interactions imex curation van-2005-1 Pam17 is required for architecture and translocation activity of the mitochondrial protein import motor. yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom ms participant Identification by mass spectrometry ms participant van der Laan M., Chacinska A., Lind M., Perschil I., Sickmann A., Meyer H.E., Guiard B., Meisinger C., Pfanner N., Rehling P. Nikolaus.Pfanner@biochemie.uni-freiburg.de, Peter.Rehling@biochemie.uni-freiburg.de Mol. Cell. Biol. (0270-7306) 2005 Only protein-protein interactions imex curation van-2005-3 Pam17 is required for architecture and translocation activity of the mitochondrial protein import motor. yeast Saccharomyces cerevisiae Baker's yeast pull down pull down western blot western blot Immuno blot Immuno blot van der Laan M., Chacinska A., Lind M., Perschil I., Sickmann A., Meyer H.E., Guiard B., Meisinger C., Pfanner N., Rehling P. Nikolaus.Pfanner@biochemie.uni-freiburg.de, Peter.Rehling@biochemie.uni-freiburg.de Mol. Cell. Biol. (0270-7306) 2005 Only protein-protein interactions imex curation tim14_yeast Mitochondrial import inner membrane translocase subunit TIM14 Presequence translocated-associated motor subunit PAM18 PAM18 TIM14 YLR008C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSQSNTGNSIEAPQLPIPGQTNGSANVTVDGAGVNVGIQNGSQGQKTGMDLYFDQALNYMGEHPVITGFGAFLTLYFTAGAYKSISKGLNGGKSTTAFLKGGFDPKMNSKEALQILNLTENTLTKKKLKEVHRKIMLANHPDKGGSPFLATKINEAKDFLEKRGISK tim50_yeast Mitochondrial import inner membrane translocase subunit TIM50 TIM50 YPL063W protein protein yeast Saccharomyces cerevisiae Baker's yeast MLSILRNSVRLNSRALRVVPSAANTLTSVQASRRLLTSYSSFLQKETKDDKPKSILTDDMLFKAGVDVDEKGQGKNEETSGEGGEDKNEPSSKSEKSRRKRQTSTDIKREKYANWFYIFSLSALTGTAIYMARDWEPQESEELKKDIDNGYTLSLMYKRFKARFNSMFTYFQEPPFPDLLPPPPPPPYQRPLTLVITLEDFLVHSEWSQKHGWRTAKRPGADYFLGYLSQYYEIVLFSSNYMMYSDKIAEKLDPIHAFVSYNLFKEHCVYKDGVHIKDLSKLNRDLSKVIIIDTDPNSYKLQPENAIPMEPWNGEADDKLVRLIPFLEYLATQQTKDVRPILNSFEDKKNLAEEFDHRVKKLKDKFYGDHKSGGNWAMTALGLGNSLGGSTKFPLDLIHEEGQKNYLMFMKMIEEEKEKIRIQQEQMGGQTFTLKDYVEGNLPSPEEQMKIQLEKQKEVDALFEEEKKKKKIAESK 778D54F5C4ED27C8 pam17_yeast Presequence translocated-associated motor subunit PAM17, mitochondrial YKR065C PAM17 FMP18 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFTSAIRLSSQRLFASQPSVTAAALRSTATTLPLRSYSQPASLQDSSILTWSDFFKLRKQQRRINVGSSLFTALLGCNVSWAYLSTMEIDPTQMLFGFDPLTVISAGIIASGALGYLLGPIVGSQVFKLSHNQQLAQFNNKNKEFLKHIINNRVDASSQSFSNPVPDYYGEKIGSLKEYKQWLRDCHAYAKKAKEFL tim23_yeast Mitochondrial import inner membrane translocase subunit TIM23 MIM23 MPI3 N3180 YNR017W TIM23 MAS6 Mitochondrial protein import protein 3 Mitochondrial protein import protein MAS6 Membrane import machinery protein MIM23 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSWLFGDKTPTDDANAAVGGQDTTKPKELSLKQSLGFEPNINNIISGPGGMHVDTARLHPLAGLDKGVEYLDLEEEQLSSLEGSQGLIPSRGWTDDLCYGTGAVYLLGLGIGGFSGMMQGLQNIPPNSPGKLQLNTVLNHITKRGPFLGNNAGILALSYNIINSTIDALRGKHDTAGSIGAGALTGALFKSSKGLKPMGYSSAMVAAACAVWCSVKKRLLEK 08E5AB4F400F675B tim17_yeast Mitochondrial import inner membrane translocase subunit TIM17 MPI2 TIM17 MIM17 SMS1 YJL143W J0648 Mitochondrial protein import protein 2 Mitochondrial inner membrane protein MIM17 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSADHSRDPCPIVILNDFGGAFAMGAIGGVVWHGIKGFRNSPLGERGSGAMSAIKARAPVLGGNFGVWGGLFSTFDCAVKAVRKREDPWNAIIAGFFTGGALAVRGGWRHTRNSSITCACLLGVIEGVGLMFQRYAAWQAKPMAPPLPEAPSSQPLQA 1760B08F3B60D378 hsp77_yeast Import motor subunit, mitochondrial ENS1 SSC1 YJR045C J1639 mtHSP70 Endonuclease SceI 75 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MLAAKNILNRSSLSSSFRIATRLQSTKVQGSVIGIDLGTTNSAVAIMEGKVPKIIENAEGSRTTPSVVAFTKEGERLVGIPAKRQAVVNPENTLFATKRLIGRRFEDAEVQRDIKQVPYKIVKHSNGDAWVEARGQTYSPAQIGGFVLNKMKETAEAYLGKPVKNAVVTVPAYFNDSQRQATKDAGQIVGLNVLRVVNEPTAAALAYGLEKSDSKVVAVFDLGGGTFDISILDIDNGVFEVKSTNGDTHLGGEDFDIYLLREIVSRFKTETGIDLENDRMAIQRIREAAEKAKIELSSTVSTEINLPFITADASGPKHINMKFSRAQFETLTAPLVKRTVDPVKKALKDAGLSTSDISEVLLVGGMSRMPKVVETVKSLFGKDPSKAVNPDEAVAIGAAVQGAVLSGEVTDVLLLDVTPLSLGIETLGGVFTRLIPRNTTIPTKKSQIFSTAAAGQTSVEIRVFQGERELVRDNKLIGNFTLAGIPPAPKGVPQIEVTFDIDADGIINVSARDKATNKDSSITVAGSSGLSENEIEQMVNDAEKFKSQDEARKQAIETANKADQLANDTENSLKEFEGKVDKAEAQKVRDQITSLKELVARVQGGEEVNAEELKTKTEELQTSSMKLFEQLYKNDSNNNNNNNGNNAESGETKQ This sequence has been withdrawn from Uniprot. 1E5672E76FCE24AC tim44_yeast Mitochondrial import inner membrane translocase subunit TIM44 ISP45 MIM44 MPI1 YIL022W TIM44 Mitochondrial protein import protein 1 Inner membrane import site protein 45 Membrane import machinery protein MIM44 protein protein yeast Saccharomyces cerevisiae Baker's yeast MHRSTFIRTSGTSSRTLTARYRSQYTGLLVARVLFSTSTTRAQGGNPRSPLQIFRDTFKKEWEKSQELQENIKTLQDASGKLGESEAYKKAREAYLKAQRGSTIVGKTLKKTGETMEHIATKAWESELGKNTRKAAAATAKKLDESFEPVRQTKIYKEVSEVIDDGESSRYGGFITKEQRRLKRERDLASGKRHRAVKSNEDAGTAVVATNIESKESFGKKVEDFKEKTVVGRSIQSLKNKLWDESENPLIVVMRKITNKVGGFFAETESSRVYSQFKLMDPTFSNESFTRHLREYIVPEILEAYVKGDVKVLKKWFSEAPFNVYAAQQKIFKEQDVYADGRILDIRGVEIVSAKLLAPQDIPVLVVGCRAQEINLYRKKKTGEIAAGDEANILMSSYAMVFTRDPEQIDDDETEGWKILEFVRGGSRQFT 8E98C738178B1E44 tim22_yeast Mitochondrial import inner membrane translocase subunit TIM22 D0884 YDL217C TIM22 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVYTGFGLEQISPAQKKPYNELTPEEQGERGAEMIMNFMTSCPGKSVVSGVTGFALGGVLGLFMASMAYDTPLHTPTPANTAATATAGNIGVGGISRTVQQISDLPFRQQMKLQFTDMGKKSYSSAKNFGYIGMIYAGVECVIESLRAKNDIYNGVTAGFFTGAGLAYKAGPQAALMGGAGFAAFSAAIDLYMKSEDGRPPQNDFKE 3C94DBF31F413968 tim54_yeast Mitochondrial import inner membrane translocase subunit TIM54 J1150 YJL054W TIM54 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSESGKPIAKPIRKPGYTNPALKALGIPALRLPSRNWMIFWSVLTVSIGGIAYDKYKQRQILSHATDLVKPLAEESMEVDKVPRKITVFIAPPPNDYLESSLKVWRRYVKPVLYYAGLDYELVQEDRQGIIRTNVANRIRELRKEILASTDGQPVKEPNQTVAKPSGSSTSKISSLLPFNKIIQDPAEEDDSFDPEIGKKFKENFDWRNVIGIFYTMPKPKHIISEDALTKDPILSGGVICLGRGAYKEYIAGIHEGLLGPIEKTEKTGSTEPKMTGVVEANQIESKVSESGATELVDAEKETALEEAKVQDDLKVDEENSSEDSQKFLKPFISSDQYPDLQIASELQTPNGEFIRNPNTNIPLLINQPLLVIPIPNLIGFTTIPRRIHRFYQKRFYVEDVCSSVVNCVRQTRIRPFDIAKDIDLAKDEEKDWPQNWVKQGKEKNSEWTQELVCDPRITKHMFVYEKPPKEEPESDI tim18_yeast Mitochondrial import inner membrane translocase subunit TIM18 TIM18 YOR297C protein protein yeast Saccharomyces cerevisiae Baker's yeast MLLFPGLKPVLNASTVIVNPVRAVFPGLVLSTKRSFYSINRLNAENKINDIANTSKEASSSVQMFKPPEFSQFKDSYQKDYERIAKYTLIPLTMVPFYASFTGGVINPLLDASLSSIFLIYLQYGFTSCIIDYIPKEKYPRWHKLALYCLYGGSMLSLYGIYELETKNNGFVDLVKKLWNENDDHLYIFGRN FE5DBEF3E3446CF7 tim21_yeast Mitochondrial import inner membrane translocase subunit TIM21 TIM21 YGR033C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSSLPRSLLRLGHRKPLFPRYNTFVNSSVITHTSLLRTRLYSNGTGATSGKKDDKTRNKPKPLWPQVKSASTFTFSGILVIGAVGISAIVIYLILSELFSPSGDTQLFNRAVSMVEKNKDIRSLLQCDDGITGKERLKAYGELITNDKWTRNRPIVSTKKLDKEGRTHHYMRFHVESKKKIALVHLEAKESKQNYQPDFINMYVDVPGEKRYYLIKPKLHPVSNSKGFLGIRWGPRKD FC140F3034E5B5BB tim16_yeast Mitochondrial import inner membrane translocase subunit TIM16 YJL104W J0822 PAM16 TIM16 Presequence translocated-associated motor subunit PAM16 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAHRAFIQVIITGTQVFGKAFAEAYRQAASQSVKQGATNASRRGTGKGEYGGITLDESCKILNIEESKGDLNMDKINNRFNYLFEVNDKEKGGSFYLQSKVYRAAERLKWELAQREKNAKAKAGDASTAKPPPNSTNSSGADNSASSNQ tom22_yeast Mitochondrial import receptor subunit TOM22 MOM22 TOM22 MAS17 MAS22 YNL131W N1217 N1862 Mitochondrial 22 kDa outer membrane protein Translocase of outer membrane 22 kDa subunit Protein MAS17 Mitochondrial 17 kDa assembly protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MVELTEIKDDVVQLDEPQFSRNQAIVEEKASATNNDVVDDEDDSDSDFEDEFDENETLLDRIVALKDIVPPGKRQTISNFFGFTSSFVRNAFTKSGNLAWTLTTTALLLGVPLSLSILAEQQLIEMEKTFDLQSDANNILAQGEKDAAATAN 6ACEC5D37D143CD3 tim23-pam16-1 1499510 n/a 1499513 unspecified role unspecified role prey prey n/a 1499514 unspecified role unspecified role prey prey n/a 1499515 unspecified role unspecified role prey prey n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499517 unspecified role unspecified role prey prey n/a 1499518 unspecified role unspecified role prey prey n/a 1499519 unspecified role unspecified role prey prey association association false false false Fig. 2A tim18-pam16 1499510 n/a 1499520 unspecified role unspecified role prey prey n/a 1499521 unspecified role unspecified role prey prey n/a 1499522 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2A tim21-tim17 1499510 n/a 1499514 unspecified role unspecified role prey prey n/a 1499517 unspecified role unspecified role prey prey n/a 1499523 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2A tim23-pam16 1499510 n/a 1499513 unspecified role unspecified role prey prey n/a 1499519 unspecified role unspecified role prey prey n/a 1499518 unspecified role unspecified role prey prey n/a 1499524 unspecified role unspecified role prey prey n/a 1499515 unspecified role unspecified role prey prey n/a 1499514 unspecified role unspecified role prey prey n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499517 unspecified role unspecified role prey prey n/a 1499523 unspecified role unspecified role prey prey association association false false false Fig. 5B tim23-pam16-2 1499510 n/a 1499519 unspecified role unspecified role prey prey n/a 1499514 unspecified role unspecified role prey prey n/a 1499518 unspecified role unspecified role prey prey n/a 1499523 unspecified role unspecified role prey prey n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499513 unspecified role unspecified role prey prey n/a 1499517 unspecified role unspecified role prey prey association association false false false Fig. 5B Interaction observed in PAM17-/- cells tim23-pam17-1 1499510 n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499519 unspecified role unspecified role prey prey n/a 1499515 unspecified role unspecified role prey prey n/a 1499523 unspecified role unspecified role prey prey n/a 1499514 unspecified role unspecified role prey prey n/a 1499513 unspecified role unspecified role prey prey association association false false false Fig. 2B tim23-pam17-2 1499510 n/a 1499523 unspecified role unspecified role prey prey n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499514 unspecified role unspecified role prey prey association association false false false Fig. 2B Interaction observed in a strain containing the temperature-sensitive mutant allele Tim17-5. tim23-pam16-3 1499511 n/a 1499523 unspecified role unspecified role prey prey n/a 1499517 unspecified role unspecified role prey prey n/a 1499519 unspecified role unspecified role prey prey n/a 1499524 unspecified role unspecified role prey prey n/a 1499518 unspecified role unspecified role prey prey n/a 1499513 unspecified role unspecified role prey prey n/a 1499516 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1499515 unspecified role unspecified role prey prey n/a 1499514 unspecified role unspecified role prey prey association association false false false Fig. 1A A contaminating fraction of the highly abundant inner membrane ADP/ATP carrier proteins was also identified. tim23-tom22 1499512 n/a 1499525 unspecified role unspecified role bait bait 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 n/a 1499514 unspecified role unspecified role prey prey n/a 1499517 unspecified role unspecified role prey prey n/a 1499519 unspecified role unspecified role prey prey association association false false false Fig. 2C When the model preprotein b2(167)delta-DHFR, consisting of a matrix-targeted amino-terminal portion of the precursor of cytochrome b2 and the passenger protein dihydrofolate reductase (DHFR), is incubated with the DHFR ligand methotrexate, the DHFR moiety cannot be unfolded by the mitochondrial import machinery. Thus, the amino-terminal portion of the preprotein accumulates in a two-membrane-spanning fashion in mitochondria, leading to the formation of a stable preprotein-TOM-TIM23-PAM supercomplex. Under these conditions TIM23 and PAM are copurified with tagged Tom22