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/ allen-2005-1 Gigaxonin-controlled degradation of MAP1B light chain is critical to neuronal survival. 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 Allen E., Ding J., Wang W., Pramanik S., Chou J., Yau V., Yang Y. Directed yeast two hybrid assay used Figure 1 (b). yyanmin@stanford.edu Nature (0028-0836) 2005 allen-2005-2 Gigaxonin-controlled degradation of MAP1B light chain is critical to neuronal survival. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Allen E., Ding J., Wang W., Pramanik S., Chou J., Yau V., Yang Y. Directed yeast two hybrid assay used Figure 1 (b). yyanmin@stanford.edu Nature (0028-0836) 2005 allen-2005-3 Gigaxonin-controlled degradation of MAP1B light chain is critical to neuronal survival. human-p12 human p12 p12 Cell line established from the peripheral blood of a 7-year-old boy with acute lymphoblastic leukemia anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Allen E., Ding J., Wang W., Pramanik S., Chou J., Yau V., Yang Y. Directed yeast two hybrid assay used Figure 1 (b). yyanmin@stanford.edu Nature (0028-0836) 2005 allen-2005-4 Gigaxonin-controlled degradation of MAP1B light chain is critical to neuronal survival. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Allen E., Ding J., Wang W., Pramanik S., Chou J., Yau V., Yang Y. Directed yeast two hybrid assay used Figure 1 (b). yyanmin@stanford.edu Nature (0028-0836) 2005 allen-2005-5 Gigaxonin-controlled degradation of MAP1B light chain is critical to neuronal survival. 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 Allen E., Ding J., Wang W., Pramanik S., Chou J., Yau V., Yang Y. Directed yeast two hybrid assay used Figure 1 (b). yyanmin@stanford.edu A human brain cDNA library was used in this study. Nature (0028-0836) 2005 gan_human Gigaxonin Kelch-like protein 16 GAN GAN1 KLHL16 protein protein human Homo sapiens Human MAEGSAVSDPQHAARLLRALSSFREESRFCDAHLVLDGEEIPVQKNILAAASPYIRTKLNYNPPKDDGSTYKIELEGISVMVMREILDYIFSGQIRLNEDTIQDVVQAADLLLLTDLKTLCCEFLEGCIAAENCIGIRDFALHYCLHHVHYLATEYLETHFRDVSSTEEFLELSPQKLKEVISLEKLNVGNERYVFEAVIRWIAHDTEIRKVHMKDVMSALWVSGLDSSYLREQMLNEPLVREIVKECSNIPLSQPQQGEAMLANFKPRGYSECIVTVGGEERVSRKPTAAMRCMCPLYDPNRQLWIELAPLSMPRINHGVLSAEGFLFVFGGQDENKQTLSSGEKYDPDANTWTALPPMNEARHNFGIVEIDGMLYILGGEDGEKELISMECYDIYSKTWTKQPDLTMVRKIGCYAAMKKKIYAMGGGSYGKLFESVECYDPRTQQWTAICPLKERRFGAVACGVAMELYVFGGVRSREDAQGSEMVTCKSEFYHDEFKRWIYLNDQNLCIPASSSFVYGAVPIGASIYVIGDLDTGTNYDYVREFKRSTGTWHHTKPLLPSDLRRTGCAALRIANCKLFRLQLQQGLFRIRVHSP uba1_human Ubiquitin-like modifier-activating enzyme 1 Ubiquitin-activating enzyme E1 Protein A1S9 UBA1 A1S9T UBE1 protein protein human Homo sapiens Human MSSSPLSKKRRVSGPDPKPGSNCSPAQSVLSEVPSVPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNTPLEDQLRVGEFCHNRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSAMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMVELNGNQPMEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLVASLAEPDFVVTDFAKFSRPAQLHIGFQALHQFCAQHGRPPRPRNEEDAAELVALAQAVNARALPAVQQNNLDEDLIRKLAYVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKEVLTEDKCLQRQNRYDGQVAVFGSDLQEKLGKQKYFLVGAGAIGCELLKNFAMIGLGCGEGGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVRQMNPHIRVTSHQNRVGPDTERIYDDDFFQNLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQRPQTWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVNNPLHLDYVMAAANLFAQTYGLTGSQDRAAVATFLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPSADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRQLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR gan-ube1 Directed yeast 2 hybrid analysis reveals interaction between the gigaxonin protein and the ubiquitin-binding domain of E1 ligase protein 256726 N/A 256731 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 256732 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The UBE1 cDNA clone was obtained from a human brain cDNA library. gan-ube1-1 Pulldown analysis reveals interaction between the gigaxonin protein and the ubiquitin-binding domain of E1 ligase protein 256727 N/A 256732 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) N/A 256731 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9H2C0:p.Val82Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false The UBE1 cDNA clone was obtained from a human brain cDNA library. The Ile 85 Phe mutation within gigaxonin reduced binding to ube1 (data not shown). gan-ube1-2 Coimmunoprecipitation analysis reveals interaction between the gigaxonin protein and the ubiquitin-binding domain of E1 ligase protein 256728 N/A 256731 unspecified role unspecified role bait bait Q9H2C0:p.Val82Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false N/A 256732 unspecified role unspecified role prey prey physical association physical association false false false gan-ube1-3 Pulldown analysis reveals interaction between the gigaxonin protein and the ubiquitin-binding domain of E1 ligase protein 256729 N/A 256732 unspecified role unspecified role prey prey human-p12 human p12 p12 Cell line established from the peripheral blood of a 7-year-old boy with acute lymphoblastic leukemia N/A 256731 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9H2C0:p.Val82Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false The UBE1 cDNA clone was obtained from a human brain cDNA library. gan-ube1-4 Directed yeast 2 hybrid analysis reveals interaction between the gigaxonin protein and the ubiquitin-binding domain of E1 ligase protein 256730 N/A 256732 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 256731 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false