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/ Interaction between amyloid precursor protein and presenilins in mammalian cells: implications for the pathogenesis of Alzheimer disease. Interaction between amyloid precursor protein and presenilins in mammalian cells: implications for the pathogenesis of Alzheimer disease. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1997 imex curation Xia W., Zhang J., Perez R., Koo EH., Selkoe DJ. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. Alzheimers - Interactions investigated in the context of Alzheimers disease selkoe@cnd.bwh.harvard.edu 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xia W., Zhang J., Perez R., Koo EH., Selkoe DJ. Apoptosis - Interactions involving proteins with a function related to apoptosis selkoe@cnd.bwh.harvard.edu Accepted 2008-NOV-10 by DAVET. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1997 davet Only protein-protein interactions imex curation Interaction between amyloid precursor protein and presenilins in mammalian cells: implications for the pathogenesis of Alzheimer disease. Interaction between amyloid precursor protein and presenilins in mammalian cells: implications for the pathogenesis of Alzheimer disease. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1997 imex curation Xia W., Zhang J., Perez R., Koo EH., Selkoe DJ. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. Alzheimers - Interactions investigated in the context of Alzheimers disease selkoe@cnd.bwh.harvard.edu 14-Nov-2008: Contacted by JYOTI. davet 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 Xia W., Zhang J., Perez R., Koo EH., Selkoe DJ. Apoptosis - Interactions involving proteins with a function related to apoptosis selkoe@cnd.bwh.harvard.edu Accepted 2008-NOV-10 by DAVET. 14-Nov-2008: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1997 davet Only protein-protein interactions imex curation o35463_crigr Amyloid-beta A4 protein beta APP protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster TRPGSGLTNIKTEEISEVKMDAEFGHDSGFEVRHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTS 37F2C6C3BFF3F597 psn2_human Presenilin-2 PSEN2 AD4 PS2 PSNL2 STM2 STM-2 E5-1 AD3LP AD5 protein protein human Homo sapiens Human MLTFMASDSEEEVCDERTSLMSAESPTPRSCQEGRQGPEDGENTAQWRSQENEEDGEEDPDRYVCSGVPGRPPGLEEELTLKYGAKHVIMLFVPVTLCMIVVVATIKSVRFYTEKNGQLIYTPFTEDTPSVGQRLLNSVLNTLIMISVIVVMTIFLVVLYKYRCYKFIHGWLIMSSLMLLFLFTYIYLGEVLKTYNVAMDYPTLLLTVWNFGAVGMVCIHWKGPLVLQQAYLIMISALMALVFIKYLPEWSAWVILGAISVYDLVAVLCPKGPLRMLVETAQERNEPIFPALIYSSAMVWTVGMAKLDPSSQGALQLPYDPEMEEDSYDSFGEPSYPEVFEPPLTGYPGEELEEEEERGVKLGLGDFIFYSVLVGKAAATGSGDWNTTLACFVAILIGLCLTLLLLAVFKKALPALPISITFGLIFYFSTDNLVRPFMDTLASHQLYI A927EEC623468116 g3hnf2_crigr Presenilin I79_012290 protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MTEIPAPLSYFQNAQMSEDSHSSSTVRSQSDSQDRQQHHDRRRLDNPEPVSNGRPQSHSRQVVEQDEEEDEELTLKYGAKHVIMLFVPVTLCMVVVVATIKSVSFYTRKDGQLIYTPFTEDTETVGQRALHSILNAAIMISVIVVMTILLVVLYKYRCYKVIHAWLIVSSLLLLFFFSFIYLGEVFKTYNVAVDYITVALLIWNFGVVGMIAIHWKGPLRLQQAYLIMISALMALVFIKYLPEWTAWLILAVISVYDLVAVLCPKGPLRMLVETAQERNETLFPALIYSSTMVWLVNMAEGDPEAQRRVPKNPKYNAQRAERERQDSSTGNDDGGFSEEWEAQRDSHLGPHRSTPESRAAVQELSSSILTNEDPEERGVKLGLGDFIFYSVLVGKASATASGDWNTTIACFVAILIGLCLTLLLLAIFKKALPALPISITFGLVFYFATDYLVQPFMDQLAFHQFYI 5F6F090E60BFFA8B 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 p05067-8 Amyloid-beta precursor protein APP A4 AD1 ABPP Alzheimer disease amyloid protein Cerebral vascular amyloid peptide PreA4 Protease nexin-II APPI PreA4 751 Amyloid precursor protein Amyloid-beta A4 protein Alzheimer disease amyloid A4 protein homolog Amyloid-beta (A4) precursor protein protein protein human Homo sapiens Human MLPGLALLLLAAWTARALEVPTDGNAGLLAEPQIAMFCGRLNMHMNVQNGKWDSDPSGTKTCIDTKEGILQYCQEVYPELQITNVVEANQPVTIQNWCKRGRKQCKTHPHFVIPYRCLVGEFVSDALLVPDKCKFLHQERMDVCETHLHWHTVAKETCSEKSTNLHDYGMLLPCGIDKFRGVEFVCCPLAEESDNVDSADAEEDDSDVWWGGADTDYADGSEDKVVEVAEEEEVAEVEEEEADDDEDDEDGDEVEEEAEEPYEEATERTTSIATTTTTTTESVEEVVREVCSEQAETGPCRAMISRWYFDVTEGKCAPFFYGGCGGNRNNFDTEEYCMAVCGSAIPTTAASTPDAVDKYLETPGDENEHAHFQKAKERLEAKHRERMSQVMREWEEAERQAKNLPKADKKAVIQHFQEKVESLEQEAANERQQLVETHMARVEAMLNDRRRLALENYITALQAVPPRPRHVFNMLKKYVRAEQKDRQHTLKHFEHVRMVDPKKAAQIRSQVMTHLRVIYERMNQSLSLLYNVPAVAEEIQDEVDELLQKEQNYSDDVLANMISEPRISYGNDALMPSLTETKTTVELLPVNGEFSLDDLQPWHSFGADSVPANTENEVEPVDARPAADRGLTTRPGSGLTNIKTEEISEVKMDAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTSIHHGVVEVDAAVTPEERHLSKMQQNGYENPTYKFFEQMQN A major isoform. C987C557C5A3714E a4_human Amyloid-beta precursor protein APP A4 AD1 Alzheimer disease amyloid protein ABPP PreA4 Cerebral vascular amyloid peptide Protease nexin-II APPI Amyloid precursor protein Amyloid-beta A4 protein Alzheimer disease amyloid A4 protein homolog Amyloid-beta (A4) precursor protein protein protein human Homo sapiens Human MLPGLALLLLAAWTARALEVPTDGNAGLLAEPQIAMFCGRLNMHMNVQNGKWDSDPSGTKTCIDTKEGILQYCQEVYPELQITNVVEANQPVTIQNWCKRGRKQCKTHPHFVIPYRCLVGEFVSDALLVPDKCKFLHQERMDVCETHLHWHTVAKETCSEKSTNLHDYGMLLPCGIDKFRGVEFVCCPLAEESDNVDSADAEEDDSDVWWGGADTDYADGSEDKVVEVAEEEEVAEVEEEEADDDEDDEDGDEVEEEAEEPYEEATERTTSIATTTTTTTESVEEVVREVCSEQAETGPCRAMISRWYFDVTEGKCAPFFYGGCGGNRNNFDTEEYCMAVCGSAMSQSLLKTTQEPLARDPVKLPTTAASTPDAVDKYLETPGDENEHAHFQKAKERLEAKHRERMSQVMREWEEAERQAKNLPKADKKAVIQHFQEKVESLEQEAANERQQLVETHMARVEAMLNDRRRLALENYITALQAVPPRPRHVFNMLKKYVRAEQKDRQHTLKHFEHVRMVDPKKAAQIRSQVMTHLRVIYERMNQSLSLLYNVPAVAEEIQDEVDELLQKEQNYSDDVLANMISEPRISYGNDALMPSLTETKTTVELLPVNGEFSLDDLQPWHSFGADSVPANTENEVEPVDARPAADRGLTTRPGSGLTNIKTEEISEVKMDAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTSIHHGVVEVDAAVTPEERHLSKMQQNGYENPTYKFFEQMQN DIP protein P05067 original sequence version: 160 DIP protein P05067 original sequence version: 207 DIP protein P05067 original sequence version: 185 DIP protein P05067 original sequence version: 188 A12EE761403740F5 psn1_human Presenilin-1 PSEN1 AD3 PS1 PSNL1 Protein S182 protein protein human Homo sapiens Human MTELPAPLSYFQNAQMSEDNHLSNTVRSQNDNRERQEHNDRRSLGHPEPLSNGRPQGNSRQVVEQDEEEDEELTLKYGAKHVIMLFVPVTLCMVVVVATIKSVSFYTRKDGQLIYTPFTEDTETVGQRALHSILNAAIMISVIVVMTILLVVLYKYRCYKVIHAWLIISSLLLLFFFSFIYLGEVFKTYNVAVDYITVALLIWNFGVVGMISIHWKGPLRLQQAYLIMISALMALVFIKYLPEWTAWLILAVISVYDLVAVLCPKGPLRMLVETAQERNETLFPALIYSSTMVWLVNMAEGDPEAQRRVSKNSKYNAESTERESQDTVAENDDGGFSEEWEAQRDSHLGPHRSTPESRAAVQELSSSILAGEDPEERGVKLGLGDFIFYSVLVGKASATASGDWNTTIACFVAILIGLCLTLLLLAIFKKALPALPISITFGLVFYFATDYLVQPFMDQLAFHQFYI DIP protein P49768 original sequence version: 106 DIP protein P49768-1 original sequence version: 144 DIP protein P49768 original sequence version: 186 5E0F451EF82BCF20 app-psen1-1 1 8 unspecified role unspecified role prey prey region (possible) glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait physical association physical association To determine whether APP and PS proteins interact in CHO cells, we performed immunoprecipitation/Western blotting using numerous distinct PS and APP antibodies. Anti-APP was specific for the human, not the hamster, protein. Fig. 2 The coprecipitating APP was almost entirely the N-glycosylated form. P49768; P05067 transfection app-psen2-1 1 8 unspecified role unspecified role prey prey region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait physical association physical association When cell lysates were precipitated with the polyclonal PS2 N-terminal antibody 2972, followed by Western blotting with APP antibody 8E5, APP was again found to coprecipitate Anti-APP was specific for the human, not the hamster, protein. Fig. 2 The coprecipitating APP was almost entirely the N-glycosylated form. P49810; P05067 transfection app-psen1-2 1 8 unspecified role unspecified role prey prey region (possible) glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait physical association physical association To determine whether APP and PS proteins interact in CHO cells, we performed immunoprecipitation/Western blotting using numerous distinct PS and APP antibodies. Anti-APP was specific for the human, not the hamster, protein. Fig. 2 The coprecipitating APP was almost entirely the N-glycosylated form. P49768; P05067 transfection app-psen1-3 1 8 unspecified role unspecified role bait bait region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey physical association physical association We also used the antibodies in reverse order: lysates were precipitated with APP antibody 8E5 and any PS1 then detected by blotting with N-terminal antibody 311/2a, �loop� antibody 4624 or C-terminal antibody 4627. Anti-APP was specific for the human, not the hamster, protein. Fig. 3B The coprecipitating APP was almost entirely the N-glycosylated form. P49768; P05067 transfection app-psen1-4 2 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region (possible) glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association P05067-4 transfection Fig. 3D N-glycosylated APP was precipitated with PS1 by antibody 4627 in 293 cells stably expressing APP695 (Fig. 3 D, lane 2). APP was N-glycosylated app-psen1-5 1 5 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To confirm unequivocally the physiological occurrence of APP-PS complexes at endogenous protein levels, we used X81 or 4627 to precipitate a large amount of lysate from nontransfected CHO cells expressing only endogenous APP and PS and then blotted with 22C11, which recognizes human and hamster APP. Fig. 3E app-psen2-2 1 4 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Using the same 22C11 antibody, endogenous hamster APP precipitated with transfected human PS2 in line C-PS2 (Fig. 3E, lane 3). Fig. 3E app-psen1-6 1 7 unspecified role unspecified role prey prey region (possible) glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined extracellular region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role bait bait physical association physical association To identify whether this region of APP is responsible for the interaction with PS, we analyzed PS1-APP complex formation in a stable CHO cell line (7WΔC) expressing APP751 truncated at residue 709 and thus lacking almost the entire cytoplasmic domain (710�751). Fig. 4 The coprecipitating APP was almost entirely the N-glycosylated form. P49768; P05067 transfection