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/ Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Biochemistry (0006-2960) 2000 imex curation Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. davet Only protein-protein interactions human-a_431 Homo sapiens epidermoid carcinoma a_431 Human epidermoid carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu Accepted 2008-MAR-12 by DAVET. 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. Biochemistry (0006-2960) 2000 davet Only protein-protein interactions imex curation Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Biochemistry (0006-2960) 2000 imex curation Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. davet 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 predetermined predetermined participant predetermined Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu Accepted 2008-MAR-12 by DAVET. 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. Biochemistry (0006-2960) 2000 davet Only protein-protein interactions imex curation Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Biochemistry (0006-2960) 2000 imex curation Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. davet Only protein-protein interactions human-a_431 Homo sapiens epidermoid carcinoma a_431 Human epidermoid carcinoma confocal microscopy confocal microscopy immunostaining immunostaining Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu Accepted 2008-MAR-12 by DAVET. 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. Biochemistry (0006-2960) 2000 davet Only protein-protein interactions imex curation Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Identification of 12-lipoxygenase interaction with cellular proteins by yeast two-hybrid screening. Biochemistry (0006-2960) 2000 imex curation Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. davet 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 Tang K., Finley RL., Nie D., Honn KV. Cancer - Interactions investigated in the context of cancer k.v.honn@wayne.edu Accepted 2008-MAR-12 by DAVET. 17-April-2008: Contacted by JYOTI. A cDNA library from human epidermoid carcinoma A431 cells. Biochemistry (0006-2960) 2000 davet Only protein-protein interactions imex curation itb4_human Integrin beta-4 GP150 ITGB4 protein protein human Homo sapiens Human MAGPRPSPWARLLLAALISVSLSGTLANRCKKAPVKSCTECVRVDKDCAYCTDEMFRDRRCNTQAELLAAGCQRESIVVMESSFQITEETQIDTTLRRSQMSPQGLRVRLRPGEERHFELEVFEPLESPVDLYILMDFSNSMSDDLDNLKKMGQNLARVLSQLTSDYTIGFGKFVDKVSVPQTDMRPEKLKEPWPNSDPPFSFKNVISLTEDVDEFRNKLQGERISGNLDAPEGGFDAILQTAVCTRDIGWRPDSTHLLVFSTESAFHYEADGANVLAGIMSRNDERCHLDTTGTYTQYRTQDYPSVPTLVRLLAKHNIIPIFAVTNYSYSYYEKLHTYFPVSSLGVLQEDSSNIVELLEEAFNRIRSNLDIRALDSPRGLRTEVTSKMFQKTRTGSFHIRRGEVGIYQVQLRALEHVDGTHVCQLPEDQKGNIHLKPSFSDGLKMDAGIICDVCTCELQKEVRSARCSFNGDFVCGQCVCSEGWSGQTCNCSTGSLSDIQPCLREGEDKPCSGRGECQCGHCVCYGEGRYEGQFCEYDNFQCPRTSGFLCNDRGRCSMGQCVCEPGWTGPSCDCPLSNATCIDSNGGICNGRGHCECGRCHCHQQSLYTDTICEINYSAIHPGLCEDLRSCVQCQAWGTGEKKGRTCEECNFKVKMVDELKRAEEVVVRCSFRDEDDDCTYSYTMEGDGAPGPNSTVLVHKKKDCPPGSFWWLIPLLLLLLPLLALLLLLCWKYCACCKACLALLPCCNRGHMVGFKEDHYMLRENLMASDHLDTPMLRSGNLKGRDVVRWKVTNNMQRPGFATHAASINPTELVPYGLSLRLARLCTENLLKPDTRECAQLRQEVEENLNEVYRQISGVHKLQQTKFRQQPNAGKKQDHTIVDTVLMAPRSAKPALLKLTEKQVEQRAFHDLKVAPGYYTLTADQDARGMVEFQEGVELVDVRVPLFIRPEDDDEKQLLVEAIDVPAGTATLGRRLVNITIIKEQARDVVSFEQPEFSVSRGDQVARIPVIRRVLDGGKSQVSYRTQDGTAQGNRDYIPVEGELLFQPGEAWKELQVKLLELQEVDSLLRGRQVRRFHVQLSNPKFGAHLGQPHSTTIIIRDPDELDRSFTSQMLSSQPPPHGDLGAPQNPNAKAAGSRKIHFNWLPPSGKPMGYRVKYWIQGDSESEAHLLDSKVPSVELTNLYPYCDYEMKVCAYGAQGEGPYSSLVSCRTHQEVPSEPGRLAFNVVSSTVTQLSWAEPAETNGEITAYEVCYGLVNDDNRPIGPMKKVLVDNPKNRMLLIENLRESQPYRYTVKARNGAGWGPEREAIINLATQPKRPMSIPIIPDIPIVDAQSGEDYDSFLMYSDDVLRSPSGSQRPSVSDDTGCGWKFEPLLGEELDLRRVTWRLPPELIPRLSASSGRSSDAEAPHGPPDDGGAGGKGGSLPRSATPGPPGEHLVNGRMDFAFPGSTNSLHRMTTTSAAAYGTHLSPHVPHRVLSTSSTLTRDYNSLTRSEHSHSTTLPRDYSTLTSVSSHDSRLTAGVPDTPTRLVFSALGPTSLRVSWQEPRCERPLQGYSVEYQLLNGGELHRLNIPNPAQTSVVVEDLLPNHSYVFRVRAQSQEGWGREREGVITIESQVHPQSPLCPLPGSAFTLSTPSAPGPLVFTALSPDSLQLSWERPRRPNGDIVGYLVTCEMAQGGGPATAFRVDGDSPESRLTVPGLSENVPYKFKVQARTTEGFGPEREGIITIESQDGGPFPQLGSRAGLFQHPLQSEYSSITTTHTSATEPFLVDGLTLGAQHLEAGGSLTRHVTQEFVSRTLTTSGTLSTHMDQQFFQT DIP protein P16144 original sequence version: 196 09710FFBBD719469 lmna_human Prelamin-A/C LMNA LMN1 protein protein human Homo sapiens Human METPSQRRATRSGAQASSTPLSPTRITRLQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSFGDNLVTRSYLLGNSSPRTQSPQNCSIM E0855F7699F0318B trib1_human Tribbles homolog 1 SKIP1 G-protein-coupled receptor-induced gene 2 protein TRIB1 C8FW GIG2 TRB1 protein protein human Homo sapiens Human MRVGPVRSAMSGASQPRGPALLFPATRGVPAKRLLDADDAAAVAAKCPRLSECSSPPDYLSPPGSPCSPQPPPAAPGAGGGSGSAPGPSRIADYLLLPLAEREHVSRALCIHTGRELRCKVFPIKHYQDKIRPYIQLPSHSNITGIVEVILGETKAYVFFEKDFGDMHSYVRSRKRLREEEAARLFKQIVSAVAHCHQSAIVLGDLKLRKFVFSTEERTQLRLESLEDTHIMKGEDDALSDKHGCPAYVSPEILNTTGTYSGKAADVWSLGVMLYTLLVGRYPFHDSDPSALFSKIRRGQFCIPEHISPKARCLIRSLLRREPSERLTAPEILLHPWFESVLEPGYIDSEIGTSDQIVPEYQEDSDISSFFC 5F54E50924B1365B lox12_human Polyunsaturated fatty acid lipoxygenase ALOX12 Arachidonate (12S)-lipoxygenase Arachidonate (15S)-lipoxygenase Linoleate (13S)-lipoxygenase ALOX12 LOG12 Platelet-type lipoxygenase 12 12LO Lipoxin synthase 12-LO protein protein human Homo sapiens Human MGRYRIRVATGAWLFSGSYNRVQLWLVGTRGEAELELQLRPARGEEEEFDHDVAEDLGLLQFVRLRKHHWLVDDAWFCDRITVQGPGACAEVAFPCYRWVQGEDILSLPEGTARLPGDNALDMFQKHREKELKDRQQIYCWATWKEGLPLTIAADRKDDLPPNMRFHEEKRLDFEWTLKAGALEMALKRVYTLLSSWNCLEDFDQIFWGQKSALAEKVRQCWQDDELFSYQFLNGANPMLLRRSTSLPSRLVLPSGMEELQAQLEKELQNGSLFEADFILLDGIPANVIRGEKQYLAAPLVMLKMEPNGKLQPMVIQIQPPNPSSPTPTLFLPSDPPLAWLLAKSWVRNSDFQLHEIQYHLLNTHLVAEVIAVATMRCLPGLHPIFKFLIPHIRYTMEINTRARTQLISDGGIFDKAVSTGGGGHVQLLRRAAAQLTYCSLCPPDDLADRGLLGLPGALYAHDALRLWEIIARYVEGIVHLFYQRDDIVKGDPELQAWCREITEVGLCQAQDRGFPVSFQSQSQLCHFLTMCVFTCTAQHAAINQGQLDWYAWVPNAPCTMRMPPPTTKEDVTMATVMGSLPDVRQACLQMAISWHLSRRQPDMVPLGHHKEKYFSGPKPKAVLNQFRTDLEKLEKEITARNEQLDWPYEYLKPSCIENSVTI C4D6D5B320666A77 k2c5_human Keratin, type II cytoskeletal 5 Cytokeratin-5 58 kDa cytokeratin KRT5 Keratin-5 Type-II keratin Kb5 protein protein human Homo sapiens Human MSRQSSVSFRSGGSRSFSTASAITPSVSRTSFTSVSRSGGGGGGGFGRVSLAGACGVGGYGSRSLYNLGGSKRISISTSGGSFRNRFGAGAGGGYGFGGGAGSGFGFGGGAGGGFGLGGGAGFGGGFGGPGFPVCPPGGIQEVTVNQSLLTPLNLQIDPSIQRVRTEEREQIKTLNNKFASFIDKVRFLEQQNKVLDTKWTLLQEQGTKTVRQNLEPLFEQYINNLRRQLDSIVGERGRLDSELRNMQDLVEDFKNKYEDEINKRTTAENEFVMLKKDVDAAYMNKVELEAKVDALMDEINFMKMFFDAELSQMQTHVSDTSVVLSMDNNRNLDLDSIIAEVKAQYEEIANRSRTEAESWYQTKYEELQQTAGRHGDDLRNTKHEISEMNRMIQRLRAEIDNVKKQCANLQNAIADAEQRGELALKDARNKLAELEEALQKAKQDMARLLREYQELMNTKLALDVEIATYRKLLEGEECRLSGEGVGPVNISVVTSSVSSGYGSGSGYGGGLGGGLGGGLGGGLAGGSSGSYYSSSSGGVGLGGGLSVGGSGFSASSGRGLGVGFGSGGGSSSSVKFVSTTSSSRKSFKS DIP protein P13647 original sequence version: 149 E9D5318E01F55145 alox12-krt5 4 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait physical association physical association we identified 12 independent clones in which the LexA-driven reporters were active only in galactose, indicating that they contained cDNA-encoded proteins that interacted with LexA-12-LOX. We isolated the library plasmids from these clones and determined that they represented four different cDNAs. Six identical clones encoded the carboxy-terminal 330 amino acid residues of type II keratin K5 alox12-lmna 4 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association we identified 12 independent clones in which the LexA-driven reporters were active only in galactose, indicating that they contained cDNA-encoded proteins that interacted with LexA-12-LOX. We isolated the library plasmids from these clones and determined that they represented four different cDNAs. Six identical clones encoded the carboxy-terminal 330 amino acid residues of type II keratin K5. Two clones encoded the carboxy-terminal 202 residues of the nuclear envelope protein lamin A. alox12-itgb4 4 8 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association we identified 12 independent clones in which the LexA-driven reporters were active only in galactose, indicating that they contained cDNA-encoded proteins that interacted with LexA-12-LOX. We isolated the library plasmids from these clones and determined that they represented four different cDNAs. Six identical clones encoded the carboxy-terminal 330 amino acid residues of type II keratin K5. Two clones encoded the carboxy-terminal 202 residues of the nuclear envelope protein lamin A. Another two clones encoded the cytoplasmic domain of integrin 4. The range of the cytoplasmic domain of integrin beta-4 has been taken from the UniProtKB entry for the protein. alox12-trib1 4 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association we identified 12 independent clones in which the LexA-driven reporters were active only in galactose, indicating that they contained cDNA-encoded proteins that interacted with LexA-12-LOX. We isolated the library plasmids from these clones and determined that they represented four different cDNAs. Six identical clones encoded the carboxy-terminal 330 amino acid residues of type II keratin K5. Two clones encoded the carboxy-terminal 202 residues of the nuclear envelope protein lamin A. Another two clones encoded the cytoplasmic domain of integrin 4. The final two clones encoded a phosphoprotein, C8FW alox12-itgb4-2 2 8 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association The range of the cytoplasmic domain of integrin beta-4 has been taken from the UniProtKB entry for the protein. Fig. 1, Table 1 alox12-krt5-1 2 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 1, Table 1 alox12-trib1-1 2 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Fig. 1, Table 1 alox12-lmna-1 2 6 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait physical association physical association Fig. 1, Table 1 alox12-krt5-2 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 2 B 12-LOX antibody co-immunoprecipitated keratin with 12-LOX in A431 cells and transfectants. alox12-krt5-3 1 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 2 B 12-LOX antibody co-immunoprecipitated keratin with 12-LOX in A431 cells and transfectants. P18054 tansfection P18054 over-expression krt5-alox12 1 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 2 B keratin antibody co-immunoprecipitated 12-LOX with keratin in A431 cells and transfectants krt5-alox12-1 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 2 B P18054 tansfection P18054 over-expression keratin antibody co-immunoprecipitated 12-LOX with keratin in A431 cells and transfectants alox12-lmna-2 1 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 4 12-LOX and lamin A coimmunoprecipitated from nuclear extracts of untransfected A431 cells. This interaction was observed in nuclear extracts and not cytoplasmic extracts. lmna-alox12 1 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association 4 12-LOX and lamin A coimmunoprecipitated from nuclear extracts of untransfected A431 cells. This interaction was observed in nuclear extracts and not cytoplasmic extracts. alox12-krt5-4 3 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 We were able to use immunofluorescent staining in A431 cells to show that 12-LOX co-localized with keratin in the cytoplasm, mostly around the cell nucleus forming a ring-like structure