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/ Preprotein translocase of the outer mitochondrial membrane: molecular dissection and assembly of the general import pore complex. Preprotein translocase of the outer mitochondrial membrane: molecular dissection and assembly of the general import pore complex. Mol. Cell. Biol. (0270-7306) 1998 Dekker PJ., Ryan MT., Brix J., Muller H., Honlinger A., Pfanner N. Mitochondria were isolated, solubilized with digitonin, and separated by blue native gel electrophoresis. After electrophoresis on a second-dimension gel under denaturing conditions, the presence of distinct Tom proteins was analyzed by immunoblotting with monospecific antisera. Since specific antibodies against Tom7 and Tom6 were not available, precursors were synthesized in vitro in rabbit reticulocyte lysates in the presence of [35S]methionine-cysteine, imported into isolated yeast mitochondria, and subjected to blue native gel electrophoresis and digital autoradiography. pfanner@ruf.uni-freiburg.de 23-JAN-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast bn-page blue native page bn-page western blot western blot Immuno blot Immuno blot Dekker PJ., Ryan MT., Brix J., Muller H., Honlinger A., Pfanner N. Mitochondria were isolated, solubilized with digitonin, and separated by blue native gel electrophoresis. After electrophoresis on a second-dimension gel under denaturing conditions, the presence of distinct Tom proteins was analyzed by immunoblotting with monospecific antisera. Since specific antibodies against Tom7 and Tom6 were not available, precursors were synthesized in vitro in rabbit reticulocyte lysates in the presence of [35S]methionine-cysteine, imported into isolated yeast mitochondria, and subjected to blue native gel electrophoresis and digital autoradiography. pfanner@ruf.uni-freiburg.de Accepted 2007-JAN-5 by DAVET. 23-JAN-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1998 Preprotein translocase of the outer mitochondrial membrane: molecular dissection and assembly of the general import pore complex. Preprotein translocase of the outer mitochondrial membrane: molecular dissection and assembly of the general import pore complex. Mol. Cell. Biol. (0270-7306) 1998 Dekker PJ., Ryan MT., Brix J., Muller H., Honlinger A., Pfanner N. Mitochondria were isolated, solubilized with digitonin, and separated by blue native gel electrophoresis. After electrophoresis on a second-dimension gel under denaturing conditions, the presence of distinct Tom proteins was analyzed by immunoblotting with monospecific antisera. Since specific antibodies against Tom7 and Tom6 were not available, precursors were synthesized in vitro in rabbit reticulocyte lysates in the presence of [35S]methionine-cysteine, imported into isolated yeast mitochondria, and subjected to blue native gel electrophoresis and digital autoradiography. pfanner@ruf.uni-freiburg.de 23-JAN-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Dekker PJ., Ryan MT., Brix J., Muller H., Honlinger A., Pfanner N. Mitochondria were isolated, solubilized with digitonin, and separated by blue native gel electrophoresis. After electrophoresis on a second-dimension gel under denaturing conditions, the presence of distinct Tom proteins was analyzed by immunoblotting with monospecific antisera. Since specific antibodies against Tom7 and Tom6 were not available, precursors were synthesized in vitro in rabbit reticulocyte lysates in the presence of [35S]methionine-cysteine, imported into isolated yeast mitochondria, and subjected to blue native gel electrophoresis and digital autoradiography. pfanner@ruf.uni-freiburg.de Accepted 2007-JAN-5 by DAVET. 23-JAN-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1998 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 6ACEC5D37D143CD3 tom6_yeast Mitochondrial import receptor subunit TOM6 YOR045W TOM6 ISP6 Mitochondrial import site protein ISP6 Translocase of outer membrane 6 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MDGMFAMPGAAAGAASPQQPKSRFQAFKESPLYTIALNGAFFVAGVAFIQSPLMDMLAPQL EE3F295BC0FE7240 EE3F295BC0FE7240 tom40_yeast Mitochondrial import receptor subunit TOM40 TOM40 ISP42 YMR203W YM8325.04 MOM38 Mitochondrial import site protein ISP42 Translocase of outer membrane 40 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAPTPLAEASQIPTIPALSPLTAKQSKGNFFSSNPISSFVVDTYKQLHSHRQSLELVNPGTVENLNKEVSRDVFLSQYFFTGLRADLNKAFSMNPAFQTSHTFSIGSQALPKYAFSALFANDNLFAQGNIDNDLSVSGRLNYGWDKKNISKVNLQISDGQPTMCQLEQDYQASDFSVNVKTLNPSFSEKGEFTGVAVASFLQSVTPQLALGLETLYSRTDGSAPGDAGVSYLTRYVSKKQDWIFSGQLQANGALIASLWRKVAQNVEAGIETTLQAGMVPITDPLMGTPIGIQPTVEGSTTIGAKYEYRQSVYRGTLDSNGKVACFLERKVLPTLSVLFCGEIDHFKNDTKIGCGLQFETAGNQELLMLQQGLDADGNPLQALPQL 6436FBBA4020464B tom20_yeast Mitochondrial import receptor subunit TOM20 MOM19 YGR082W TOM20 MAS20 Mitochondrial 20 kDa outer membrane protein Translocase of outer membrane 20 kDa subunit Protein MAS20 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQSNPILRGLAITTAIAALSATGYAIYFDYQRRNSPQFRKVLRQRAKEQAKMEEQAKTHAKEVKLQKVTEFLSMELAKDPIPSDPSEREATFTTNVENGERLSMQQGKELEAASKFYKALTVYPQPADLLGIYQRSIPEAIYEYIILMIAILPPANVASFVKGVVGSKAESDAVAEANDIDD 9F76FE6010278410 tom7_yeast Mitochondrial import receptor subunit TOM7 TOM7 MOM7 YNL070W N2378 Translocase of outer membrane 7 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MSFLPSFILSDESKERISKILTLTHNVAHYGWIPFVLYLGWAHTSNRPNFLNLLSPLPSV 0EFADB326E8926E3 tom5_yeast Mitochondrial import receptor subunit TOM5 YPR133W-A TOM5 MOM8A Translocase of outer membrane 5 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MFGLPQQEVSEEEKRAHQEQTEKTLKQAAYVAAFLWVSPMIWHLVKKQWK E1419ED5FEC45E14 E1419ED5FEC45E14 tom6-tom40-3 1 6 unspecified role unspecified role neutral component neutral component 8 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 7 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 3 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 5 unspecified role unspecified role neutral component neutral component 4 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 Molecular weight = 450kDa 1 Upon overexposure of the immunoblot, a small amount of Tom20(~5 to 10%) was found in the higher-molecular-weight range, slightly above the peak of the 400K position (small amounts of Tom40 and other components of the GIP complex were also observed at this slightly higher position) tom6-tom40-4 1 7 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 3 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 5 unspecified role unspecified role neutral component neutral component 8 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 4 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 Molecular weight = 400kDa 1 We conclude that Tom40, Tom22, and the three small Tom proteins comigrate at the 400K position in blue native gel electrophoresis. tom20-tom40-3 2 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait association association 2A Coimmunoprecipitation of Tom70, Tom20, and Tom22 from digitonin-lysed mitochondria was performed with all available monospecific anti-Tom antibodies: anti-Tom70, anti-Tom20, anti-Tom22, anti-Tom40, and anti-Tom5. tom20-tom22-1 2 6 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey physical association physical association 2A Coimmunoprecipitation of Tom70, Tom20, and Tom22 from digitonin-lysed mitochondria was performed with all available monospecific anti-Tom antibodies: anti-Tom70, anti-Tom20, anti-Tom22, anti-Tom40, and anti-Tom5. tom20-tom22-2 2 6 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait physical association physical association 2A Coimmunoprecipitation of Tom70, Tom20, and Tom22 from digitonin-lysed mitochondria was performed with all available monospecific anti-Tom antibodies: anti-Tom70, anti-Tom20, anti-Tom22, anti-Tom40, and anti-Tom5. tom20-tom22-3 2 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 3 unspecified role unspecified role prey prey association association 2A Coimmunoprecipitation of Tom70, Tom20, and Tom22 from digitonin-lysed mitochondria was performed with all available monospecific anti-Tom antibodies: anti-Tom70, anti-Tom20, anti-Tom22, anti-Tom40, and anti-Tom5.