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/ keskanokwon-2007-3 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-4 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. pull down pull down anti tag western anti tag western blot anti tag western Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. Co2+ chelate resin used to pull down bait proteins. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-1 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. 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 Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-2 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. 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 predetermined predetermined participant predetermined Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-5 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. His-tagged ILK and kAE1 were co-expressed in HEK 293 cells, and radiolabeled with L-[35S]methionine for 1 h, followed by a 24-h chase period. His-tagged ILK and any associated proteins were purified by nickel-affinity resin. Eluted associated proteins were then incubated with anti-Ct AE1 to immunoprecipitate kAE1 and with anti-ILK to immunoprecipitate ILK. Autoradiographs revealed the interaction of ILK and kAE1. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-6 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. canlf-mdck Canis familiaris (dog) kidney epithelial cells Canis lupus familiaris Dog Canis familiaris mdck Canine Cocker Spaniel kidney epithelial cell line confocal microscopy confocal microscopy immunostaining immunostaining Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation keskanokwon-2007-7 Interaction of integrin-linked kinase with the kidney chloride/bicarbonate exchanger, kAE1. human-kidney Human kidney kidney kidney [Renal] confocal microscopy confocal microscopy immunostaining immunostaining Keskanokwong T., Shandro HJ., Johnson DE., Kittanakom S., Vilas GL., Thorner P., Reithmeier RA., Akkarapatumwong V., Yenchitsomanus PT., Casey JR. joe.casey@ualberta.ca Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation ilk_human Integrin-linked protein kinase ILK-1 ILK-2 59 kDa serine/threonine-protein kinase p59ILK ILK ILK1 ILK2 Beta-integrin-linked kinase protein protein human Homo sapiens Human MDDIFTQCREGNAVAVRLWLDNTENDLNQGDDHGFSPLHWACREGRSAVVEMLIMRGARINVMNRGDDTPLHLAASHGHRDIVQKLLQYKADINAVNEHGNVPLHYACFWGQDQVAEDLVANGALVSICNKYGEMPVDKAKAPLRELLRERAEKMGQNLNRIPYKDTFWKGTTRTRPRNGTLNKHSGIDFKQLNFLTKLNENHSGELWKGRWQGNDIVVKVLKVRDWSTRKSRDFNEECPRLRIFSHPNVLPVLGACQSPPAPHPTLITHWMPYGSLYNVLHEGTNFVVDQSQAVKFALDMARGMAFLHTLEPLIPRHALNSRSVMIDEDMTARISMADVKFSFQCPGRMYAPAWVAPEALQKKPEDTNRRSADMWSFAVLLWELVTREVPFADLSNMEIGMKVALEGLRPTIPPGISPHVCKLMKICMNEDPAKRPKFDMIVPILEKMQDK nadap_human Kanadaptin SLC4A1AP Kidney anion exchanger adapter protein Solute carrier family 4 anion exchanger member 1 adapter protein Human lung cancer oncogene 3 protein HLC3 protein protein human Homo sapiens Human MLAPLRNAPGREGATSPSPPTDATGSLGEWDVDRNVKTEGWVSKERISKLHRLRMADILSQSETLASQDLSGDFKKPALPVSPAARSKAPASSSSNPEEVQKEGPTALQDSNSGEPDIPPPQPDCGDFRSLQEEQSRPPTAVSSPGGPARAPPYQEPPWGGPATAPYSLETLKGGTILGTRSLKGTSYCLFGRLSGCDVCLEHPSVSRYHAVLQHRASGPDGECDSNGPGFYLYDLGSTHGTFLNKTRIPPRTYCRVHVGHVVRFGGSTRLFILQGPEEDREAESELTVTQLKELRKQQQILLEKKMLGEDSDEEEEMDTSERKINAGSQDDEMGCTWGMGEDAVEDDAEENPIVLEFQQEREAFYIKDPKKALQGFFDREGEELEYEFDEQGHSTWLCRVRLPVDDSTGKQLVAEAIHSGKKKEAMIQCSLEACRILDTLGLLRQEAVSRKRKAKNWEDEDFYDSDDDTFLDRTGLIEKKRLNRMKKAGKIDEKPETFESLVAKLNDAERELSEISERLKASSQVLSESPSQDSLDAFMSEMKSGSTLDGVSRKKLHLRTFELRKEQQRLKGLIKIVKPAEIPELKKTETQTTGAENKAKKLTLPLFGAMKGGSKFKLKTGTVGKLPPKRPELPPTLMRMKDEPEVEEEEEEEEEEEKEKEEHEKKKLEDGSLSRPQPEIEPEAAVQEMRPPTDLTHFKETQTHENMSQLSEEEQNKDYQDCSKTTSLCAGPSASKNEYEKSRGELKKKKTPGPGKLPPTLSSKYPEDDPDYCVWVPPEGQSGDGRTHLNDKYGY parva_human Alpha-parvin Calponin-like integrin-linked kinase-binding protein CH-ILKBP Matrix-remodeling-associated protein 2 Actopaxin PARVA MXRA2 protein protein human Homo sapiens Human MATSPQKSPSVPKSPTPKSPPSRKKDDSFLGKLGGTLARRKKAKEVSELQEEGMNAINLPLSPIPFELDPEDTMLEENEVRTMVDPNSRSDPKLQELMKVLIDWINDVLVGERIIVKDLAEDLYDGQVLQKLFEKLESEKLNVAEVTQSEIAQKQKLQTVLEKINETLKLPPRSIKWNVDSVHAKSLVAILHLLVALSQYFRAPIRLPDHVSIQVVVVQKREGILQSRQIQEEITGNTEALSGRHERDAFDTLFDHAPDKLNVVKKTLITFVNKHLNKLNLEVTELETQFADGVYLVLLMGLLEGYFVPLHSFFLTPDSFEQKVLNVSFAFELMQDGGLEKPKPRPEDIVNCDLKSTLRVLYNLFTKYRNVE paxi_human Paxillin PXN protein protein human Homo sapiens Human MDDLDALLADLESTTSHISKRPVFLSEETPYSYPTGNHTYQEIAVPPPVPPPPSSEALNGTILDPLDQWQPSSSRFIHQQPQSSSPVYGSSAKTSSVSNPQDSVGSPCSRVGEEEHVYSFPNKQKSAEPSPTVMSTSLGSNLSELDRLLLELNAVQHNPPGFPADEANSSPPLPGALSPLYGVPETNSPLGGKAGPLTKEKPKRNGGRGLEDVRPSVESLLDELESSVPSPVPAITVNQGEMSSPQRVTSTQQQTRISASSATRELDELMASLSDFKIQGLEQRADGERCWAAGWPRDGGRSSPGGQDEGGFMAQGKTGSSSPPGGPPKPGSQLDSMLGSLQSDLNKLGVATVAKGVCGACKKPIAGQVVTAMGKTWHPEHFVCTHCQEEIGSRNFFERDGQPYCEKDYHNLFSPRCYYCNGPILDKVVTALDRTWHPEHFFCAQCGAFFGPEGFHEKDGKAYCRKDYFDMFAPKCGGCARAILENYISALNTLWHPECFVCRECFTPFVNGSFFEHDGQPYCEVHYHERRGSLCSGCQKPITGRCITAMAKKFHPEHFVCAFCLKQLNKGTFKEQNDKPYCQNCFLKLFC e2r5h4_canlf 59 kDa serine/threonine-protein kinase ILK Beta-integrin-linked kinase ILK-1 ILK-2 Integrin-linked protein kinase p59ILK protein protein canlf Canis familiaris (dog) Canis lupus familiaris Dog Canis familiaris MDDIFTQCREGNAVAVRLWLDNTENDLNQGDDHGFSPLHWACREGRSAVVEMLIMRGARINVMNRGDDTPLHLAASHGHRDIVQKLLQYKADINAVNEHGNVPLHYACFWGQDQVAEDLVANGALVSICNKYGEMPVDKAKAPLRELLRERAEKMGQNLNRIPYKDTFWKGTTRTRPRNGTLNKHSGIDFKQLNFLAKLNENHSGELWKGRWQGNDIVVKVLKVRDWSTRKSRDFNEECPRLRIFSHPNVLPVLGACQSPPAPHPTLITHWMPYGSLYNVLHEGTNFVVDQSQAVKFALDMARGMAFLHTLEPLIPRHALNSRSVMIDEDMTARISMADVKFSFQCPGRMYAPAWVAPEALQKKPEDTNRRSADMWSFAVLLWELVTREVPFADLSNMEIGMKVALEGLRPTIPPGISPHVCKLMKICMNEDPAKRPKFDMIVPILEKMQDK Similar to Q13418 slc4a1ap-ilk-5 Interaction between SLC4A1AP and ILK demonstrated by coimmunoprecipitation 768717 n/a 768724 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 768725 unspecified role unspecified role prey prey c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Figs. 1 and 2 An active kinase domain of ILK1 is not required for SLC4A1AP binding slc4a1ap-ilk-6 Interaction between SLC4A1AP and ILK demonstrated by coimmunoprecipitation 768717 n/a 768724 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 768725 unspecified role unspecified role prey prey c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ch domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 1, Fig. 4 slc4a1ap-ilk-4 Interaction between SLC4A1AP and ILK demonstrated by pulldown 768718 n/a 768725 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 768724 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig. 1 slc4a1ap-ilk-8 Interaction between SLC4A1AP and ILK demonstrated by pulldown 768718 n/a 768725 unspecified role unspecified role prey prey n/a 768726 unspecified role unspecified role prey prey n/a 768724 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 768727 unspecified role unspecified role prey prey physical association physical association false false false Fig. 8 Paxillin or actopaxin were not detected when kAE1 was expressed without ILK suggesting that kAE1 interacts with the actin cytoskeleton through a complex containing ILK, paxillin, and actopaxin. slc4a1ap-ilk-1 Interaction between SLC4A1AP and ILK demonstrated by Y2H 768719 n/a 768724 unspecified role unspecified role prey prey n/a 768725 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false slc4a1ap-ilk-2 Interaction between SLC4A1AP and ILK demonstrated by Y2H 768720 n/a 768724 unspecified role unspecified role prey prey n/a 768725 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Table 1 slc4a1ap-ilk-7 Interaction between SLC4A1AP and ILK demonstrated by coimmunoprecipitation 768721 n/a 768725 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 768724 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 7 slc4a1ap-ilk-9 Colocalisation of SL4A1Ap and ILK 768722 n/a 768725 unspecified role unspecified role neutral component neutral component c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 768728 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 10 Colocalisation only observed in polarised cells slc4a1ap-ilk-10 Colocalisation of SL4A1Ap and ILK 768723 n/a 768724 unspecified role unspecified role neutral component neutral component n/a 768725 unspecified role unspecified role neutral component neutral component c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig. 11