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/ Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions mouse-lymphoblast Mus musculus lymphoblast Transformed lymphocytes; Blast cell; Lymphoblastoid solution sedimentati cosedimentation in solution solution sedimentati western blot western blot Immuno blot Immuno blot Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. The authors have mentioned lymphocytic cells as being the host cells. please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions mouse-lymphoblast Mus musculus lymphoblast Transformed lymphocytes; Blast cell; Lymphoblastoid fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. The authors have mentioned T cells as being the host cells. please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions mouse-baf Mus musculus pro-B cells baf mouse pro-B cell line cannot establish identity to CABRI: ECACC 87033001 since furher info is missing. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. please notice that in the description of mutations by author substitute by with of and of with by. Analysis was carried out in cells expressing the constitutively active Rap1V12. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat. Immunol. (1529-2908) 2006 imex curation Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp 02-MAR-2006: Contacted by JYOTI. human cDNA library Only protein-protein interactions 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Katagiri K., Imamura M., Kinashi T. Synapse - Interactions of proteins with an established role in the presynapse. please notice that in the description of mutations by author substitute by with of and of with by. kinashi@takii.kmu.ac.jp Accepted 2007-FEB-5 by DAVET. 02-MAR-2006: Contacted by JYOTI. human cDNA library Nat. Immunol. (1529-2908) 2006 Only protein-protein interactions imex curation itb2_mouse Integrin beta-2 Cell surface adhesion glycoproteins LFA-1/CR3/p150,95 subunit beta Complement receptor C3 subunit beta Itgb2 protein protein mouse Mus musculus Mouse MLGLRPSLLLALAGLFFLGSAVSQECTKYKVSSCRDCIQSGPGCSWCQKLNFTGPGEPDSLRCDTRAQLLLKGCPADDIMDPRSIANPEFDQRGQRKQLSPQKVTLYLRPGQAAAFNVTFRRAKGYPIDLYYLMDLSYSMLDDLNNVKKLGGDLLQALNEITESGRIGFGSFVDKTVLPFVNTHPEKLRNPCPNKEKACQPPFAFRHVLKLTDNSNQFQTEVGKQLISGNLDAPEGGLDAIMQVAACPEEIGWRNVTRLLVFATDDGFHFAGDGKLGAILTPNDGRCHLEDNMYKRSNEFDYPSVGQLAHKLSESNIQPIFAVTKKMVKTYEKLTEIIPKSAVGELSDDSSNVVQLIKNAYYKLSSRVFLDHSTLPDTLKVTYDSFCSNGASSIGKSRGDCDGVQINNPVTFQVKVMASECIQEQSFVIRALGFTDTVTVQVRPQCECQCRDQSREQSLCGGKGVMECGICRCESGYIGKNCECQTQGRSSQELERNCRKDNSSIVCSGLGDCICGQCVCHTSDVPNKEIFGQYCECDNVNCERYNSQVCGGSDRGSCNCGKCSCKPGYEGSACQCQRSTTGCLNARLVECSGRGHCQCNRCICDEGYQPPMCEDCPSCGSHCRDNHTSCAECLKFDKGPFEKNCSVQCAGMTLQTIPLKKKPCKERDSEGCWITYTLQQKDGRNIYNIHVEDSLECVKGPNVAAIVGGTVVGVVLIGVLLLVIWKALTHLTDLREYRRFEKEKLKSQWNNDNPLFKSATTTVMNPKFAES DIP protein P11835 original sequence version: 95 6FA51DA93AB40D8A q8www0-2 Ras association domain-containing protein 5 B RASSF5 NORE1 RAPL New ras effector 1 Regulator for cell adhesion and polarization enriched in lymphoid tissues protein protein human Homo sapiens Human MTVDSSMSSGYCSLDEELEDCFFTAKTTFFRNAQSKHLSKNVCKPVEETQRPPTLQEIKQKIDSYNTREKNCLGMKLSEDGTYTGFIKVHLKLRRPVTVPAGIRPQSIYDAIKEVNLAATTDKRTSFYLPLDAIKQLHISSTTTVSEVIQGLLKKFMVVDNPQKFALFKRIHKDGQVLFQKLSIADRPLYLRLLAGPDTEVLSFVLKENETGEVEWDAFSIPELQNFLTILEKEEQDKIQQVQKKYDKFRQKLEEALRESQGKPG Initiator Met-1 is removed. Contains a N-acetylthreonine at position 2. 66AD5D84EFA6D6CB stk3_human Serine/threonine-protein kinase 3 STE20-like kinase MST2 Mammalian STE20-like protein kinase 2 Serine/threonine-protein kinase Krs-1 STK3 MST2 KRS1 protein protein human Homo sapiens Human MEQPPAPKSKLKKLSEDSLTKQPEEVFDVLEKLGEGSYGSVFKAIHKESGQVVAIKQVPVESDLQEIIKEISIMQQCDSPYVVKYYGSYFKNTDLWIVMEYCGAGSVSDIIRLRNKTLIEDEIATILKSTLKGLEYLHFMRKIHRDIKAGNILLNTEGHAKLADFGVAGQLTDTMAKRNTVIGTPFWMAPEVIQEIGYNCVADIWSLGITSIEMAEGKPPYADIHPMRAIFMIPTNPPPTFRKPELWSDDFTDFVKKCLVKNPEQRATATQLLQHPFIKNAKPVSILRDLITEAMEIKAKRHEEQQRELEEEEENSDEDELDSHTMVKTSVESVGTMRATSTMSEGAQTMIEHNSTMLESDLGTMVINSEDEEEEDGTMKRNATSPQVQRPSFMDYFDKQDFKNKSHENCNQNMHEPFPMSKNVFPDNWKVPQDGDFDFLKNLSLEELQMRLKALDPMMEREIEELRQRYTAKRQPILDAMDAKKRRQQNF DIP protein Q13188 original sequence version: 150 DIP protein Q13188 original sequence version: 139 84F598ED3617292C rasf5_mouse Ras association domain-containing protein 5 Rassf5 Nore1 Rapl New ras effector 1 Regulator for cell adhesion and polarization enriched in lymphoid tissues protein protein mouse Mus musculus Mouse MASPAIGQRPYPLLLDPEPPRYLQSLGGTEPPPPARPRRCIPTALIPAAGASEDRGGRRSGRRDPEPTPRDCRHARPVRPGLQPRLRLRPGSHRPRDVRSIFEQPQDPRVLAERGEGHRFVELALRGGPGWCDLCGREVLRQALRCANCKFTCHSECRSLIQLDCRQKGGPALDRRSPESTLTPTLNQNVCKAVEETQHPPTIQEIKQKIDSYNSREKHCLGMKLSEDGTYTGFIKVHLKLRRPVTVPAGIRPQSIYDAIKEVNPAATTDKRTSFYLPLDAIKQLHISSTTTVSEVIQGLLKKFMVVDNPQKFALFKRIHKDGQVLFQKLSIADYPLYLRLLAGPDTDVLSFVLKENETGEVEWDAFSIPELQNFLTILEKEEQDKIHQLQKKYNKFRQKLEEALRESQGKPG DIP protein Q5EBH1 original sequence version: 22 E6FF7DDE6BECB180 stk4_mouse Serine/threonine-protein kinase 4 STE20-like kinase MST1 Mammalian STE20-like protein kinase 1 Stk4 Mst1 protein protein mouse Mus musculus Mouse METVQLRNPPRRQLKKLDEDSLTKQPEEVFDVLEKLGEGSYGSVYKAIHKETGQIVAIKQVPVESDLQEIIKEISIMQQCDSPHVVKYYGSYFKNTDLWIVMEYCGAGSVSDIIRLRNKTLTEDEIATILQSTLKGLEYLHFMRKIHRDIKAGNILLNTEGHAKLADFGVAGQLTDTMAKRNTVIGTPFWMAPEVIQEIGYNCVADIWSLGITAIEMAEGKPPYADIHPMRAIFMIPTNPPPTFRKPELWSDNFMDFVKQCLVKSPEQRATATQLLQHPFVKSAKGVSILRDLINEAMDVKLKRQEAQQREVDQDDEENSEEDEMDSGTMVRAAGDEMGTVRVASTMSGGANTMIEHGDTLPSQLGTMVINTEDEEEEGTMKRRDETMQPAKPSFLEYFEQKEKENQINSFGKNVSGSLKNSSDWKIPQDGDYEFLKSWTVEDLQKRLLALDPMMEQEMEEIRQKYRSKRQPILDAIEAKKRRQQNF A21E9519AD209C40 eea1_mouse Early endosome antigen 1 Eea1 protein protein mouse Mus musculus Mouse MFRRILQRTPGRVGSQGSDLDSSATPINTVDVNNESSSEGFICPQCMKSLGSADELFKHYQAVHDAGNDSGHGGEAGLALTRDDITLLRQEVQDLQASLKEEKWYSEELKKELEKYQGLQQQEAKSDGLVTDSSAELQALEQQLEEAQTENFNIKQMKDLFEQKAAQLATEIADIKSKYDEEKSLRAAAEQKVTHLTEDLNKQTTVIQDLKTELLQRPGIEDVAVLKKELVQVQTLMDNMTLERERESEKLKDECKKLQSEHAHLEATINQLRSELAKGPQEVAVYVQEIQKLKGSINELTQKNQNLTEKLQKKDLDYTHLEEKHNEESASRKTLQASLHQRDLDCQQLQARLTASESSLQRAQGELSEKAEAAQKLREELREVESTRQHLKVEVKQLQQQREEKEQHGLQLQGEVSQLHCKLLETERQLGEAHGRLKEQRQLSSEKLMEKEQQVADLQLKLSRLEEQLKEKVTNSTELQHQLEKSKQQHQEQQALQQSATAKLREAQNDLEQVLRQIGDKDQKIQNLEALLQKGKESVSLLEKEREDLYAKIQAGEGETAVLNQLQEKNHALQQQLTQLTEKLKNQSESHKQAEENLHDQVQEQKAHLRAAQDRVLSLETSVSELSSQLNESKEKVSQLDIQIKAKTELLLSAEAAKAAQRADLQNHLDTAQHALQDKQQELNKVSVQLDQLTAKFQEKQEHCIQLESHLKDHKEKHLSLEQKVEDLEGHIKKLEADALEVKASKEQALQSLQQQRQLSTDLELRNAELSRELQEQEEVVSCTKLDLQNKSEILENIKQTLTKKEEENVVLKQEFEKLSQDSKTQHKELGDRMQAAVTELTAVKAQKDALLAELSTTKEKLSKVSDSLKNSKSEFEKENQKGKAAVLDLEKACKELKHQLQVQAESALKEQEDLKKSLEKEKETSQQLKIELNSVKGEVSQAQNTLKQKEKDEQQLQGTINQLKQSAEQKKKQIEALQGEVKNAVSQKTVLENKLQQQSSQAAQELAAEKGKLSALQSNYEKCQADLKQLQSDLYGKESELLATRQDLKSVEEKLTLAQEDLISNRNQIGNQNKSIQELQAAKASLEQDSAKKEALLKEQSKALEDAQREKSVKEKELVAEKSKLAEMEEIKCRQEKEITKLNEELKSHKQESIKEITNLKDAKQLLIQQKLELQGRVDSLKAALEQEKESQQLMREQVKKEEEKRKEEFSEKEAKLHSEIKEKEAGMKKHEENEAKLTMQVTTLNENLGTVKKEWQSSQRRVSELEKQTDDLRGEIAVLEATVQNNQDERRALLERCLKGEGEIEKLQTKALELQRKLDNTTAAVQELGRENQSLQIKHTQALNRKWAEDNEVQNCMSCGKCFSVTVRRHHCRQCGNIFCAECSTKNALTPSSKKPVRVCDACFNDLQG 2365A51EF92019FD stk4_human Serine/threonine-protein kinase 4 STK4 MST1 STE20-like kinase MST1 Mammalian STE20-like protein kinase 1 Serine/threonine-protein kinase Krs-2 KRS2 protein protein human Homo sapiens Human METVQLRNPPRRQLKKLDEDSLTKQPEEVFDVLEKLGEGSYGSVYKAIHKETGQIVAIKQVPVESDLQEIIKEISIMQQCDSPHVVKYYGSYFKNTDLWIVMEYCGAGSVSDIIRLRNKTLTEDEIATILQSTLKGLEYLHFMRKIHRDIKAGNILLNTEGHAKLADFGVAGQLTDTMAKRNTVIGTPFWMAPEVIQEIGYNCVADIWSLGITAIEMAEGKPPYADIHPMRAIFMIPTNPPPTFRKPELWSDNFTDFVKQCLVKSPEQRATATQLLQHPFVRSAKGVSILRDLINEAMDVKLKRQESQQREVDQDDEENSEEDEMDSGTMVRAVGDEMGTVRVASTMTDGANTMIEHDDTLPSQLGTMVINAEDEEEEGTMKRRDETMQPAKPSFLEYFEQKEKENQINSFGKSVPGPLKNSSDWKIPQDGDYEFLKSWTVEDLQKRLLALDPMMEQEIEEIRQKYQSKRQPILDAIEAKKRRQQNF 150758EBC5F77D5C rassf5-mst1 6 8 unspecified role unspecified role bait bait Q8WWW0-2:p.[Lys123Ala;Arg124Ala;Lys135Ala;Lys154Ala;Lys155Ala;Asp160Ala;Asn161Ala] lys 123, 135, 154, 155; arg 124; asp 160; asn 161 replaced by ala mutation mutation certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain D A Fri Jul 22 11:10:30 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg124ala,lys154ala,lys123ala,asn161ala,lys155ala,lys135ala,asp160ala' Wed May 02 16:45:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys123ala,arg124ala,lys135ala,lys154ala,lys155ala,asp160ala,asn161ala' Tue Jun 30 18:01:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys123Ala;Arg124Ala;Lys135Ala;Lys154Ala;Lys155Ala;Asp160Ala;Asn161Ala]' 13 unspecified role unspecified role prey prey physical association physical association Using an activated RAPL mutant as the bait, we isolated cDNA encoding human MST1 or MST2 by yeast two-hybrid screening An activated form of RAPL (M4) was generated by the substitution by alanine of lysine positions 123, 135, 154 and 155, arginine at position 124, aspartic acid at position 160, and asparagine at position 161. rassf5-mst2 6 8 unspecified role unspecified role bait bait Q8WWW0-2:p.[Lys123Ala;Arg124Ala;Lys135Ala;Lys154Ala;Lys155Ala;Asp160Ala;Asn161Ala] lys 123, 135, 154, 155; arg 124; asp 160; asn 161 replaced by ala mutation mutation certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain D A Fri Jul 22 11:07:07 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys154ala,arg124ala,asn161ala,lys123ala,lys135ala,lys155ala,asp160ala' Wed May 02 15:24:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys123ala,arg124ala,lys135ala,lys154ala,lys155ala,asp160ala,asn161ala' Tue Jun 30 16:48:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys123Ala;Arg124Ala;Lys135Ala;Lys154Ala;Lys155Ala;Asp160Ala;Asn161Ala]' 9 unspecified role unspecified role prey prey physical association physical association Using an activated RAPL mutant as the bait, we isolated cDNA encoding human MST1 or MST2 by yeast two-hybrid screening RAPL (M4) was generated by the introduction of substitution by alanine of lysine residues at positions 123, 135, 154 and 155, of arginine at position 124, of aspartic acid at position 160, and of asparagine at position 161. rassf5-mst1-1 3 8 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role prey prey region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association MST1 and RAPL could be immunoprecipitated together using antibody to V5 (anti-V5) or anti-Myc. Deletion of the C-terminal region of RAPL abolished its association with MST1 but deletion of the N-terminal region of RAPL did not (Fig. 1a) 1a rassf5-mst1-2 5 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined coiled coil region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role prey prey c terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 physical association physical association 1b A fusion protein of glutathione S-transferase (GST) and the RAPL coiled-coil domain precipitated wildtype MST1 but not the MST1 mutant lacking the C-terminal regulatory region (Fig. 1b). These results indicate that the coiled-coil region of RAPL associates with the MST1 regulatory domain. rassf5-mst1-3 4 10 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association 1c in the mouse pro�B cell line BAF stably expressing a constitutively active mutant form of Rap1 (Rap1V12) or vector alone (as a control). RAPL and Mst1 were associated each other in the absence of exogenous stimuli. Interaction was carried out in cells expressing the constitutively active Rap1V12. rassf5-mst1-4 4 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association 1c in the mouse pro�B cell line BAF stably expressing a constitutively active mutant form of Rap1 (Rap1V12) or vector alone (as a control). RAPL and Mst1 were associated each other in the absence of exogenous stimuli. Interaction was carried out in the cells expressing the constitutively active Rap1V12. rassf5-mst1-5 3 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait c terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association MST1 and RAPL could be immunoprecipitated together using antibody to V5 (anti-V5) or anti-Myc. Deletion of the C-terminal region of RAPL abolished its association with MST1 but deletion of the N-terminal region of RAPL did not (Fig. 1a) 1a rassf5-mst1-6 2 10 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component physical association physical association 2e RAPL and Mst1 localized together in unstimulated T cells and also at the leading edge in CCL21-stimulated T cells RAPL and Mst1 localized together in unstimulated T cells and also at the leading edge in CCL21-stimulated T cells mst1-rassf5 1 10 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component physical association physical association 7b Rab5, Rab11, and Rap1 also formed part of this complex however since the subspecies of protein involved could not be identified UniProtKB accession numbers could not be assigned. Subcellular fractionation by continuous sucrose density gradient centrifugation showed that RAPL and Mst1 fractionated together exclusively in light fractions (fractions 2 and 3), which contained early (EEA1and Rab5) and late (Rab11) endosomal marker proteins, but lacked beta-COP and transferrin receptor, indicative of the Golgi-derived vesicles and recycling endosomes, respectively (Fig. 7b). An complex was also observed between proteins TfR, Rap1 and beta2 integrin however, the UniProtKB accession numbers for TfR and Rap1 coudl not be determined since the subforms were not specified.