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/ The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. J. Biol. Chem. (0021-9258) 2005 imex curation Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Apoptosis - Interactions involving proteins with a function related to apoptosis Alzheimers - Interactions investigated in the context of Alzheimers disease Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu 27-Oct-2007: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Apoptosis - Interactions involving proteins with a function related to apoptosis Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu Accepted 2007-MAY-23 by ORCHARD. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. J. Biol. Chem. (0021-9258) 2005 imex curation Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Apoptosis - Interactions involving proteins with a function related to apoptosis Alzheimers - Interactions investigated in the context of Alzheimers disease Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu 27-Oct-2007: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Apoptosis - Interactions involving proteins with a function related to apoptosis Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu Accepted 2007-MAY-23 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production. J. Biol. Chem. (0021-9258) 2005 imex curation Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Apoptosis - Interactions involving proteins with a function related to apoptosis Alzheimers - Interactions investigated in the context of Alzheimers disease Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu 27-Oct-2007: Contacted by JYOTI. Only protein-protein interactions human-brain Homo sapiens brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Matsuda S., Giliberto L., Matsuda Y., Davies P., McGowan E., Pickford F., Ghiso J., Frangione B., D'Adamio L. Apoptosis - Interactions involving proteins with a function related to apoptosis Authors also state that yeast 2 hybrid screening of a human brain cDNA library resulted in identification of two proteins (ITM2A and ITM2B) that interact with APP family members but do not stipulate whether the interaction is with APP or APLP2. Thus the interactions could not be annotated. HeLa cells co-transfected with ITM2A and APP constructs, cell lysates immunoprecipated with aFLAG antibody. ldadamio@aecom.yu.edu Accepted 2007-MAY-23 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation p05067-4 Amyloid-beta precursor protein APP A4 AD1 ABPP Alzheimer disease amyloid protein Cerebral vascular amyloid peptide PreA4 Protease nexin-II APPI PreA4 695 Amyloid precursor protein Amyloid-beta A4 protein Alzheimer disease amyloid A4 protein homolog Amyloid-beta (A4) precursor protein protein protein human Homo sapiens Human MLPGLALLLLAAWTARALEVPTDGNAGLLAEPQIAMFCGRLNMHMNVQNGKWDSDPSGTKTCIDTKEGILQYCQEVYPELQITNVVEANQPVTIQNWCKRGRKQCKTHPHFVIPYRCLVGEFVSDALLVPDKCKFLHQERMDVCETHLHWHTVAKETCSEKSTNLHDYGMLLPCGIDKFRGVEFVCCPLAEESDNVDSADAEEDDSDVWWGGADTDYADGSEDKVVEVAEEEEVAEVEEEEADDDEDDEDGDEVEEEAEEPYEEATERTTSIATTTTTTTESVEEVVRVPTTAASTPDAVDKYLETPGDENEHAHFQKAKERLEAKHRERMSQVMREWEEAERQAKNLPKADKKAVIQHFQEKVESLEQEAANERQQLVETHMARVEAMLNDRRRLALENYITALQAVPPRPRHVFNMLKKYVRAEQKDRQHTLKHFEHVRMVDPKKAAQIRSQVMTHLRVIYERMNQSLSLLYNVPAVAEEIQDEVDELLQKEQNYSDDVLANMISEPRISYGNDALMPSLTETKTTVELLPVNGEFSLDDLQPWHSFGADSVPANTENEVEPVDARPAADRGLTTRPGSGLTNIKTEEISEVKMDAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTSIHHGVVEVDAAVTPEERHLSKMQQNGYENPTYKFFEQMQN DIP protein P05067-4 original sequence version: 183 DIP protein P05067-4 original sequence version: 198 DIP protein P05067-4 original sequence version: 224 A major isoform. 4F6EA0139F969D56 itm2a_human Integral membrane protein 2A ITM2A UNQ603/PRO1189 Protein E25 protein protein human Homo sapiens Human MVKIAFNTPTAVQKEEARQDVEALLSRTVRTQILTGKELRVATQEKEGSSGRCMLTLLGLSFILAGLIVGGACIYKYFMPKSTIYRGEMCFFDSEDPANSLRGGEPNFLPVTEEADIREDDNIAIIDVPVPSFSDSDPAAIIHDFEKGMTAYLDLLLGNCYLMPLNTSIVMPPKNLVELFGKLASGRYLPQTYVVREDLVAVEEIRDVSNLGIFIYQLCNNRKSFRLRRRDLLLGFNKRAIDKCWKIRHFPNEFIVETKICQE C565E2F80FE223BD aplp1-itm2a 2 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminal region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig 1E APP mouse monoclonal antibody 22C11 (Chemicon) was used to pull down APP interactions. A 17kDa proteolytic N terminal fragment also binds APP. itm2a-aplp1 3 5 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 2b C83 isofirm did not precipate with ITM2A. EN3 anti ITM2A rabbit polyclonal antibody raised against aa 222-232 of human sequence used to immunoprecipate ITM2A complexes. itm2a-aplp1-1 1 5 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 n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figs. 1b-d The C83 isoform of APP was not found to bind to ITM2A Immunoprecipitation of cell lysates with an {alpha}FLAG antibody showed that BRI2 interacted with full-length A{beta}PP (Fig. 1, b and d), C99 (Fig. 1, b and d), and A{beta}PPNcas, which present a deletion of most of the intracellular region of A{beta}PP (Fig. 1c) but not C83 (Fig. 1, b and d).