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/ Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. J. Biol. Chem. (0021-9258) 2004 Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de 02-MAR-2006: Contacted by JYOTI. Returned mail. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de Accepted 2007-FEB-15 by JYOTI. 02-MAR-2006: Contacted by JYOTI. Returned mail. J. Biol. Chem. (0021-9258) 2004 Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. J. Biol. Chem. (0021-9258) 2004 Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de 02-MAR-2006: Contacted by JYOTI. Returned mail. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de Accepted 2007-FEB-15 by JYOTI. 02-MAR-2006: Contacted by JYOTI. Returned mail. J. Biol. Chem. (0021-9258) 2004 Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. Organization and function of the small Tim complexes acting along the import pathway of metabolite carriers into mammalian mitochondria. J. Biol. Chem. (0021-9258) 2004 Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de 02-MAR-2006: Contacted by JYOTI. Returned mail. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Muhlenbein N., Hofmann S., Rothbauer U., Bauer MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Mitochondrion lysed with 0.5% digitonin Mitochondrion lysed with 0.5% digitonin bauer@bio.med.uni-muenchen.de Accepted 2007-FEB-15 by JYOTI. 02-MAR-2006: Contacted by JYOTI. Returned mail. J. Biol. Chem. (0021-9258) 2004 tim9_human Mitochondrial import inner membrane translocase subunit Tim9 TIMM9 TIM9 TIM9A TIMM9A protein protein human Homo sapiens Human MAAQIPESDQIKQFKEFLGTYNKLTETCFLDCVKDFTTREVKPEETTCSEHCLQKYLKMTQRISMRFQEYHIQQNEALAAKAGLLGQPR 00F17CF6332ABF48 tim10_mouse Mitochondrial import inner membrane translocase subunit Tim10 Timm10 Tim10 protein protein mouse Mus musculus Mouse MDPLRAQQLAAELEVEMMADMYNRMTSACHRKCVPPHYKEAELSKGESVCLDRCVSKYLDIHERMGKKLTELSMQDEELMKRVQQSSGPA D20EFAE694D14BAB adt3_human ADP/ATP translocase 3 SLC25A6 ANT3 CDABP0051 Adenine nucleotide translocator 3 ADP,ATP carrier protein 3 Solute carrier family 25 member 6 ADP,ATP carrier protein, isoform T2 AAC3 protein protein human Homo sapiens Human MTEQAISFAKDFLAGGIAAAISKTAVAPIERVKLLLQVQHASKQIAADKQYKGIVDCIVRIPKEQGVLSFWRGNLANVIRYFPTQALNFAFKDKYKQIFLGGVDKHTQFWRYFAGNLASGGAAGATSLCFVYPLDFARTRLAADVGKSGTEREFRGLGDCLVKITKSDGIRGLYQGFSVSVQGIIIYRAAYFGVYDTAKGMLPDPKNTHIVVSWMIAQTVTAVAGVVSYPFDTVRRRMMMQSGRKGADIMYTGTVDCWRKIFRDEGGKAFFKGAWSNVLRGMGGAFVLVLYDELKKVI 18534E9F0E49672F tim22_human Mitochondrial import inner membrane translocase subunit Tim22 TIMM22 TEX4 TIM22 Testis-expressed protein 4 protein protein human Homo sapiens Human MAAAAPNAGGSAPETAGSAEAPLQYSLLLQYLVGDKRQPRLLEPGSLGGIPSPAKSEEQKMIEKAMESCAFKAALACVGGFVLGGAFGVFTAGIDTNVGFDPKDPYRTPTAKEVLKDMGQRGMSYAKNFAIVGAMFSCTECLIESYRGTSDWKNSVISGCITGGAIGFRAGLKAGAIGCGGFAAFSAAIDYYLR A1E6DD9F7F7399D6 t10b_human Mitochondrial import inner membrane translocase subunit Tim10 B TIMM10B FXC1 Fracture callus protein 1 FxC1 Mitochondrial import inner membrane translocase subunit Tim9 B TIM9B TIMM10B TIMM9B protein protein human Homo sapiens Human MERQQQQQQQLRNLRDFLLVYNRMTELCFQRCVPSLHHRALDAEEEACLHSCAGKLIHSNHRLMAAYVQLMPALVQRRIADYEAASAVPGVAAEQPGVSPSGS B76CCC81668B1F31 tim10_human Mitochondrial import inner membrane translocase subunit Tim10 TIMM10 TIM10 protein protein human Homo sapiens Human MDPLRAQQLAAELEVEMMADMYNRMTSACHRKCVPPHYKEAELSKGESVCLDRCVSKYLDIHERMGKKLTELSMQDEELMKRVQQSSGPA D20EFAE694D14BAB tim9_mouse Mitochondrial import inner membrane translocase subunit Tim9 Timm9 Tim9 Tim9a Timm9a protein protein mouse Mus musculus Mouse MAAQIPESDQIKQFKEFLGTYNKLTETCFLDCVKDFTTREVKPEEVTCSEHCLQKYLKMTQRISVRFQEYHIQQNEALAAKAGLLGQPR 1A40D7F491A09548 timm9-timm10-1 1 8 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey association association Immunoblot analysis of precipitated material showed that antibodies against Tim9 precipitated both Tim9 and Tim10a; likewise, both proteins were precipitated using antibodies against Tim10a indicating that both proteins interact in a complex (Fig. 2). 2 timm9-timm10-2 1 8 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Immunoblot analysis of precipitated material showed that antibodies against Tim9 precipitated both Tim9 and Tim10a; likewise, both proteins were precipitated using antibodies against Tim10a indicating that both proteins interact in a complex (Fig. 2). 2 timm9-timm10-3 1 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association Immunoblot analysis of precipitated material showed that antibodies against Tim9 precipitated both Tim9 and Tim10a; likewise, both proteins were precipitated using antibodies against Tim10a indicating that both proteins interact in a complex (Fig. 2). 2 timm9-timm10-4 2 4 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component physical association physical association Native complex sizes of Tim9, Tim10a, and Tim10b. HeLa cell mitochondria were solubilized in 0.5% digitonin (A and B, left panels) or in 0.5% Triton X-100 (A and B, right panels) and subjected to gel filtration on a Superose 12 column. 3 Molecular weight = 450kDa timm9-timm10-5 2 4 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component physical association physical association Native complex sizes of Tim9, Tim10a, and Tim10b. HeLa cell mitochondria were solubilized in 0.5% digitonin (A and B, left panels) or in 0.5% Triton X-100 (A and B, right panels) and subjected to gel filtration on a Superose 12 column. 3 Molecular weight = 70kDa timm9-timm10-6 1 4 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Western blot analysis of the immunoprecipitated material showed that both anti-Tim9 and anti-Tim10b were able to co-precipitate Tim22 4 timm9-timm10-7 1 8 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Western blot analysis of the immunoprecipitated material showed that both anti-Tim9 and anti-Tim10b were able to co-precipitate Tim22 (Fig. 4). 4 timm9-timm10-8 1 7 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Western blot analysis of the immunoprecipitated material showed that both anti-Tim9 and anti-Tim10b were able to co-precipitate Tim22 (Fig. 4). 4 timm9-slc25a6-1 3 6 unspecified role unspecified role neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 10 unspecified role unspecified role neutral component neutral component mouse-liver Mus musculus liver liver physical association physical association To address whether the human small Tims directly interact with translocation intermediates of carriers during import into mammalian mitochondria, human ANT3 was subjected to chemical cross-linking. 5 timm10-slc25a6-2 3 5 unspecified role unspecified role neutral component neutral component mouse-liver Mus musculus liver liver 6 unspecified role unspecified role neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association To address whether the human small Tims directly interact with translocation intermediates of carriers during import into mammalian mitochondria, human ANT3 was subjected to chemical cross-linking. 5