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/ Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Oncogene (0950-9232) 2006 imex curation Verissimo F., Silva E., Morris JD., Pepperkok R., Jordan P. Apoptosis - Interactions involving proteins with a function related to apoptosis HSP-70 also identified as interacting with WNK3 but Ab does not discriminate between HSPA1A or HSPA2 Antibody array specific to 400 cellular proteins involved in signal transduction (www.hypromatrix.com). The array was incubated with an extract from Myc-WNK3-expressing HEK293 cells that was prepared under non-denaturing conditions to preserve protein-protein interactions. Cellular proteins were allowed to bind to their cognate antibodies on the array membrane and Myc-WNK3 interaction then stained by chemiluminescence with an anti-Myc peroxidase conjugate. peter.jordan@insa.min-saude.pt Only protein-protein interactions 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. antibody array antibody array sandwich immunoassay antigen capture assay antibody detection identification by antibody antibody detection Verissimo F., Silva E., Morris JD., Pepperkok R., Jordan P. Apoptosis - Interactions involving proteins with a function related to apoptosis HSP-70 also identified as interacting with WNK3 but Ab does not discriminate between HSPA1A or HSPA2 Antibody array specific to 400 cellular proteins involved in signal transduction (www.hypromatrix.com). The array was incubated with an extract from Myc-WNK3-expressing HEK293 cells that was prepared under non-denaturing conditions to preserve protein-protein interactions. Cellular proteins were allowed to bind to their cognate antibodies on the array membrane and Myc-WNK3 interaction then stained by chemiluminescence with an anti-Myc peroxidase conjugate. peter.jordan@insa.min-saude.pt Accepted 2007-JAN-16 by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Oncogene (0950-9232) 2006 imex curation Verissimo F., Silva E., Morris JD., Pepperkok R., Jordan P. Apoptosis - Interactions involving proteins with a function related to apoptosis HSP-70 also identified as interacting with WNK3 but Ab does not discriminate between HSPA1A or HSPA2 Antibody array specific to 400 cellular proteins involved in signal transduction (www.hypromatrix.com). The array was incubated with an extract from Myc-WNK3-expressing HEK293 cells that was prepared under non-denaturing conditions to preserve protein-protein interactions. Cellular proteins were allowed to bind to their cognate antibodies on the array membrane and Myc-WNK3 interaction then stained by chemiluminescence with an anti-Myc peroxidase conjugate. peter.jordan@insa.min-saude.pt Only protein-protein interactions 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Verissimo F., Silva E., Morris JD., Pepperkok R., Jordan P. Apoptosis - Interactions involving proteins with a function related to apoptosis HSP-70 also identified as interacting with WNK3 but Ab does not discriminate between HSPA1A or HSPA2 Antibody array specific to 400 cellular proteins involved in signal transduction (www.hypromatrix.com). The array was incubated with an extract from Myc-WNK3-expressing HEK293 cells that was prepared under non-denaturing conditions to preserve protein-protein interactions. Cellular proteins were allowed to bind to their cognate antibodies on the array membrane and Myc-WNK3 interaction then stained by chemiluminescence with an anti-Myc peroxidase conjugate. peter.jordan@insa.min-saude.pt Accepted 2007-JAN-16 by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation wnk3_human Serine/threonine-protein kinase WNK3 Protein kinase with no lysine 3 Protein kinase lysine-deficient 3 WNK3 KIAA1566 PRKWNK3 protein protein human Homo sapiens Human MATDSGDPASTEDSEKPDGISFENRVPQVAATLTVEARLKEKNSTFSASGETVERKRFFRKSVEMTEDDKVAESSPKDERIKAAMNIPRVDKLPSNVLRGGQEVKYEQCSKSTSEISKDCFKEKNEKEMEEEAEMKAVATSPSGRFLKFDIELGRGAFKTVYKGLDTETWVEVAWCELQDRKLTKAEQQRFKEEAEMLKGLQHPNIVRFYDSWESILKGKKCIVLVTELMTSGTLKTYLKRFKVMKPKVLRSWCRQILKGLQFLHTRTPPIIHRDLKCDNIFITGPTGSVKIGDLGLATLMRTSFAKSVIGTPEFMAPEMYEEHYDESVDVYAFGMCMLEMATSEYPYSECQNAAQIYRKVTSGIKPASFNKVTDPEVKEIIEGCIRQNKSERLSIRDLLNHAFFAEDTGLRVELAEEDDCSNSSLALRLWVEDPKKLKGKHKDNEAIEFSFNLETDTPEEVAYEMVKSGFFHESDSKAVAKSIRDRVTPIKKTREKKPAGCLEERRDSQCKSMGNVFPQPQNTTLPLAPAQQTGAECEETEVDQHVRQQLLQRKPQQHCSSVTGDNLSEAGAASVIHSDTSSQPSVAYSSNQTMGSQMVSNIPQAEVNVPGQIYSSQQLVGHYQQVSGLQKHSKLTQPQILPLVQGQSTVLPVHVLGPTVVSQPQVSPLTVQKVPQIKPVSQPVGAEQQAALLKPDLVRSLNQDVATTKENVSSPDNPSGNGKQDRIKQRRASCPRPEKGTKFQLTVLQVSTSGDNMVECQLETHNNKMVTFKFDVDGDAPEDIADYMVEDNFVLESEKEKFVEELRAIVGQAQEILHVHFATERATGVDSITVDSNSSQTGSSEQVQINSTSTQTSNESAPQSSPVGRWRFCINQTIRNRETQSPPSLQHSMSAVPGRHPLPSPKNTSNKEISRDTLLTIENNPCHRALFTSKSEHKDVVDGKISECASVETKQPAILYQVEDNRQIMAPVTNSSSYSTTSVRAVPAECEGLTKQASIFIPVYPCHQTASQADALMSHPGESTQTSGNSLTTLAFDQKPQTLSVQQPAMDAEFISQEGETTVNTEASSPKTVIPTQTPGLEPTTLQPTTVLESDGERPPKLEFADNRIKTLDEKLRNLLYQEHSISSIYPESQKDTQSIDSPFSSSAEDTLSCPVTEVIAISHCGIKDSPVQSPNFQQTGSKLLSNVAASQPANISVFKRDLNVITSVPSELCLHEMSSDASLPGDPEAYPAAVSSGGAIHLQTGGGYFGLSFTCPSLKNPISKKSWTRKLKSWAYRLRQSTSFFKRSKVRQVETEEMRSAIAPDPIPLTRESTADTRALNRCKAMSGSFQRGRFQVITIPQQQSAKMTSFGIEHISVFSETNHSSEEAFIKTAKSQLVEIEPATQNPKTSFSYEKLQALQETCKENKGVPKQGDNFLSFSAACETDVSSVTPEKEFEETSATGSSMQSGSELLLKEREILTAGKQPSSDSEFSASLAGSGKSVAKTGPESNQCLPHHEEQAYAQTQSSLFYSPSSPMSSDDESEIEDEDLKVELQRLREKHIQEVVNLQTQQNKELQELYERLRSIKDSKTQSTEIPLPPASPRRPRSFKSKLRSRPQSLTHVDNGIVATGKSCLINELENPLCVESNAASCQQSPASKKGMFTDDLHKLVDDWTKEAVGNSLIKPSLNQLKQSQHKLETENWNKVSENTPSTMGYTSTWISSLSQIRGAVPTSLPQGLSLPSFPGPLSSYGMPHVCQYNAVAGAGYPVQWVGISGTTQQSVVIPAQSGGPFQPGMNMQAFPTSSVQNPATIPPGPK 1766D6AC8C7DE864 casp3_human Caspase-3 Apopain Cysteine protease CPP32 Protein Yama SREBP cleavage activity 1 CASP3 CPP32 protein protein human Homo sapiens Human MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIIINNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTAPGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESFSFDATFHAKKQIPCIVSMLTKELYFYH DIP protein P42574 original sequence version: 108 DIP protein P42574 original sequence version: 171 DIP protein P42574 original sequence version: 111 DIP protein P42574 original sequence version: 105 DIP protein P42574 original sequence version: 131 DIP protein P42574 original sequence version: 132 DIP protein P42574 original sequence version: 134 DIP protein P42574 original sequence version: 162 DIP protein P42574 original sequence version: 189 DIP protein P42574 original sequence version: 167 2F35CD3BCF7FF64A wnk3-casp3-1 1 3 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 4 unspecified role unspecified role bait bait physical association physical association WNK3 interacts with proteins involved in the regulation of apoptosis 5A wnk3-casp3-2 2 4 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association A commercial antiprocaspase-3 antibody was used to precipitate procaspase-3 and the precipitate was found to contain GFP-WNK3 (Figure 5b) 5B Interaction independent of kinase actvity of WNK3