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/ blaser-2006-1 Human endothelial cell septins: SEPT11 is an interaction partner of SEPT5. 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 Blaser S., Roseler S., Rempp H., Bartsch I., Bauer H., Lieber M., Lessmann E., Weingarten L., Busse A., Huber M., Zieger B. Synapse - Interactions of proteins with an established role in the presynapse. SEPT5 was coupled to CNBr-activated Sepharose 4B. zieger@kikli.ukl.uni-freiburg.de Human brain cDNA library. Human adult heart cDNA library. Human fetal brain cDNA library. J. Pathol. (0022-3417) 2006 Only protein-protein interactions imex curation blaser-2006-2 Human endothelial cell septins: SEPT11 is an interaction partner of SEPT5. 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 Blaser S., Roseler S., Rempp H., Bartsch I., Bauer H., Lieber M., Lessmann E., Weingarten L., Busse A., Huber M., Zieger B. Synapse - Interactions of proteins with an established role in the presynapse. SEPT5 was coupled to CNBr-activated Sepharose 4B. zieger@kikli.ukl.uni-freiburg.de Human brain cDNA library. Human adult heart cDNA library. Human fetal brain cDNA library. J. Pathol. (0022-3417) 2006 Only protein-protein interactions imex curation blaser-2006-3 Human endothelial cell septins: SEPT11 is an interaction partner of SEPT5. in vitro In vitro affinity techniques affinity technology affinity techniques western blot western blot Immuno blot Immuno blot Blaser S., Roseler S., Rempp H., Bartsch I., Bauer H., Lieber M., Lessmann E., Weingarten L., Busse A., Huber M., Zieger B. Synapse - Interactions of proteins with an established role in the presynapse. SEPT5 was coupled to CNBr-activated Sepharose 4B. zieger@kikli.ukl.uni-freiburg.de Human brain cDNA library. Human adult heart cDNA library. Human fetal brain cDNA library. J. Pathol. (0022-3417) 2006 Only protein-protein interactions imex curation blaser-2006-4 Human endothelial cell septins: SEPT11 is an interaction partner of SEPT5. chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Blaser S., Roseler S., Rempp H., Bartsch I., Bauer H., Lieber M., Lessmann E., Weingarten L., Busse A., Huber M., Zieger B. Synapse - Interactions of proteins with an established role in the presynapse. SEPT5 was coupled to CNBr-activated Sepharose 4B. zieger@kikli.ukl.uni-freiburg.de Human brain cDNA library. Human adult heart cDNA library. Human fetal brain cDNA library. J. Pathol. (0022-3417) 2006 Only protein-protein interactions imex curation sep11_human Septin-11 SEPTIN11 SEPT11 protein protein human Homo sapiens Human MAVAVGRPSNEELRNLSLSGHVGFDSLPDQLVNKSTSQGFCFNILCVGETGIGKSTLMDTLFNTKFESDPATHNEPGVRLKARSYELQESNVRLKLTIVDTVGFGDQINKDDSYKPIVEYIDAQFEAYLQEELKIKRSLFNYHDTRIHACLYFIAPTGHSLKSLDLVTMKKLDSKVNIIPIIAKADTIAKNELHKFKSKIMSELVSNGVQIYQFPTDEETVAEINATMSVHLPFAVVGSTEEVKIGNKMAKARQYPWGVVQVENENHCDFVKLREMLIRVNMEDLREQTHTRHYELYRRCKLEEMGFKDTDPDSKPFSLQETYEAKRNEFLGELQKKEEEMRQMFVMRVKEKEAELKEAEKELHEKFDLLKRTHQEEKKKVEDKKKELEEEVNNFQKKKAAAQLLQSQAQQSGAQQTKKDKDKKNASFT D467F1F315FFD028 sept5_human Septin-5 SEPTIN5 Cell division control-related protein 1 PNUTL1 Peanut-like protein 1 SEPT5 protein protein human Homo sapiens Human MSTGLRYKSKLATPEDKQDIDKQYVGFATLPNQVHRKSVKKGFDFTLMVAGESGLGKSTLVHSLFLTDLYKDRKLLSAEERISQTVEILKHTVDIEEKGVKLKLTIVDTPGFGDAVNNTECWKPITDYVDQQFEQYFRDESGLNRKNIQDNRVHCCLYFISPFGHGLRPVDVGFMKALHEKVNIVPLIAKADCLVPSEIRKLKERIREEIDKFGIHVYQFPECDSDEDEDFKQQDRELKESAPFAVIGSNTVVEAKGQRVRGRLYPWGIVEVENQAHCDFVKLRNMLIRTHMHDLKDVTCDVHYENYRAHCIQQMTSKLTQDSRMESPIPILPLPTPDAETEKLIRMKDEELRRMQEMLQRMKQQMQDQ 47054765DEA10D33 sept8_human Septin-8 SEPTIN8 KIAA0202 SEPT8 protein protein human Homo sapiens Human MAATDLERFSNAEPEPRSLSLGGHVGFDSLPDQLVSKSVTQGFSFNILCVGETGIGKSTLMNTLFNTTFETEEASHHEACVRLRPQTYDLQESNVQLKLTIVDAVGFGDQINKDESYRPIVDYIDAQFENYLQEELKIRRSLFDYHDTRIHVCLYFITPTGHSLKSLDLVTMKKLDSKVNIIPIIAKADTISKSELHKFKIKIMGELVSNGVQIYQFPTDDEAVAEINAVMNAHLPFAVVGSTEEVKVGNKLVRARQYPWGVVQVENENHCDFVKLREMLIRVNMEDLREQTHSRHYELYRRCKLEEMGFQDSDGDSQPFSLQETYEAKRKEFLSELQRKEEEMRQMFVNKVKETELELKEKERELHEKFEHLKRVHQEEKRKVEEKRRELEEETNAFNRRKAAVEALQSQALHATSQQPLRKDKDKKNRSDIGAHQPGMSLSSSKVMMTKASVEPLNCSSWWPAIQCCSCLVRDATWREGFL sept5-sept11 Yeast 2 hybrid analysis reveals interaction between SEPT5 protein and SEPT11 protein 491309 N/A 491313 unspecified role unspecified role prey prey N/A 491314 unspecified role unspecified role bait bait physical association physical association false false false SEPT5 was found to interact with SEPT11 in human brain, human fetal brain and human heart cDNA libraries. sept5-sept8 Yeast 2 hybrid analysis reveals interaction between SEPT5 protein and SEPT8 protein 491309 N/A 491314 unspecified role unspecified role bait bait N/A 491315 unspecified role unspecified role prey prey physical association physical association false false false SEPT5 was found to interact with SEPT8 in human brain and human fetal brain cDNA libraries. sept11-sept5 Yeast 2 hybrid analysis reveals interaction between SEPT11 protein and SEPT5 protein 491309 N/A 491314 unspecified role unspecified role prey prey N/A 491313 unspecified role unspecified role bait bait physical association physical association false false false SEPT11 was found to interact with SEPT5 human heart cDNA library but not in human brain cDNA library. sept5-sept11-1 Directed yeast 2 hybrid analysis reveals interaction between SEPT5 protein and SEPT11 protein 491310 N/A 491313 unspecified role unspecified role prey prey N/A 491314 unspecified role unspecified role bait bait Q99719:p.[Gly56del;Lys57del;Ser58Asn] 56Gly del 57 Lys del Ser58Asn mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 Figure 1 C. sept5-sept8-1 Directed yeast 2 hybrid analysis reveals interaction between SEPT5 protein and SEPT8 protein 491310 N/A 491314 unspecified role unspecified role bait bait N/A 491315 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 1 A. sept11-sept5-1 Directed yeast 2 hybrid analysis reveals interaction between SEPT11 protein and SEPT5 protein 491310 N/A 491313 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 Q9NVA2:p.Ser55Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false N/A 491314 unspecified role unspecified role prey prey physical association physical association false false false Figure 1 B. sept5-sept11-2 Affinity chromatography analysis reveals interaction between SEPT5 protein and SEPT11 protein 491311 N/A 491313 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia N/A 491314 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 2. sept5-sept11-3 Fret analysis reveals interaction between SEPT5 protein and SEPT11 protein 491312 N/A 491314 unspecified role unspecified role fluorescence donor fluorescence donor n terminal enhanced cyan fluorescence protein tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 491313 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept n terminal enhanced yellow fluorscent protein tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3