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/ Binding of F-spondin to amyloid-beta precursor protein: a candidate amyloid-beta precursor protein ligand that modulates amyloid-beta precursor protein cleavage. Binding of F-spondin to amyloid-beta precursor protein: a candidate amyloid-beta precursor protein ligand that modulates amyloid-beta precursor protein cleavage. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2004 imex curation Ho A., Sudhof TC. 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 Amino acid numbering in publication refers to APP695 isoform and thus differs from the numbering used in this entry. GST and Ig fusion proteins used to pull down interacting proteins from membrane proteins solublised from rat brain or COS cells. thomas.sudhof@utsouthwestern.edu 27-Oct-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ho A., Sudhof TC. Apoptosis - Interactions involving proteins with a function related to apoptosis Amino acid numbering in publication refers to APP695 isoform and thus differs from the numbering used in this entry. GST and Ig fusion proteins used to pull down interacting proteins from membrane proteins solublised from rat brain or COS cells. thomas.sudhof@utsouthwestern.edu Accepted 2007-MAY-22 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2004 Only protein-protein interactions imex curation Binding of F-spondin to amyloid-beta precursor protein: a candidate amyloid-beta precursor protein ligand that modulates amyloid-beta precursor protein cleavage. Binding of F-spondin to amyloid-beta precursor protein: a candidate amyloid-beta precursor protein ligand that modulates amyloid-beta precursor protein cleavage. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2004 imex curation Ho A., Sudhof TC. 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 Amino acid numbering in publication refers to APP695 isoform and thus differs from the numbering used in this entry. GST and Ig fusion proteins used to pull down interacting proteins from membrane proteins solublised from rat brain or COS cells. thomas.sudhof@utsouthwestern.edu 27-Oct-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Ho A., Sudhof TC. Apoptosis - Interactions involving proteins with a function related to apoptosis Amino acid numbering in publication refers to APP695 isoform and thus differs from the numbering used in this entry. GST and Ig fusion proteins used to pull down interacting proteins from membrane proteins solublised from rat brain or COS cells. thomas.sudhof@utsouthwestern.edu Accepted 2007-MAY-22 by ORCHARD. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2004 Only protein-protein interactions imex curation aplp2_human Amyloid beta precursor like protein 2 APPH APPL2 APLP2 Amyloid protein homolog CDEI box-binding protein Amyloid beta (A4) precursor-like protein 2 Amyloid-like protein 2 Sperm membrane protein YWK-II protein protein human Homo sapiens Human MAATGTAAAAATGRLLLLLLVGLTAPALALAGYIEALAANAGTGFAVAEPQIAMFCGKLNMHVNIQTGKWEPDPTGTKSCFETKEEVLQYCQEMYPELQITNVMEANQRVSIDNWCRRDKKQCKSRFVTPFKCLVGEFVSDVLLVPEKCQFFHKERMEVCENHQHWHTVVKEACLTQGMTLYSYGMLLPCGVDQFHGTEYVCCPQTKIIGSVSKEEEEEDEEEEEEEDEEEDYDVYKSEFPTEADLEDFTEAAVDEDDEDEEEGEEVVEDRDYYYDTFKGDDYNEENPTEPGSDGTMSDKEITHDVKAVCSQEAMTGPCRAVMPRWYFDLSKGKCVRFIYGGCGGNRNNFESEDYCMAVCKAMIPPTPLPTNDVDVYFETSADDNEHARFQKAKEQLEIRHRNRMDRVKKEWEEAELQAKNLPKAERQTLIQHFQAMVKALEKEAASEKQQLVETHLARVEAMLNDRRRMALENYLAALQSDPPRPHRILQALRRYVRAENKDRLHTIRHYQHVLAVDPEKAAQMKSQVMTHLHVIEERRNQSLSLLYKVPYVAQEIQEEIDELLQEQRADMDQFTASISETPVDVRVSSEESEEIPPFHPFHPFPALPENEDTQPELYHPMKKGSGVGEQDGGLIGAEEKVINSKNKVDENMVIDETLDVKEMIFNAERVGGLEEERESVGPLREDFSLSSSALIGLLVIAVAIATVIVISLVMLRKRQYGTISHGIVEVDPMLTPEERHLNKMQNHGYENPTYKYLEQMQI DIP protein Q06481 original sequence version: 94 CA3A7D6DDB8A28D0 spon1_human Spondin-1 VSGP SPON1 KIAA0762 F-spondin Vascular smooth muscle cell growth-promoting factor protein protein human Homo sapiens Human MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYTEFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLNVRAAPSAEFSVDRTRHLMSFLTMMGPSPDWNVGLSAEDLCTKECGWVQKVVQDLIPWDAGTDSGVTYESPNKPTIPQEKIRPLTSLDHPQSPFYDPEGGSITQVARVVIERIARKGEQCNIVPDNVDDIVADLAPEEKDEDDTPETCIYSNWSPWSACSSSTCDKGKRMRQRMLKAQLDLSVPCPDTQDFQPCMGPGCSDEDGSTCTMSEWITWSPCSISCGMGMRSRERYVKQFPEDGSVCTLPTEETEKCTVNEECSPSSCLMTEWGEWDECSATCGMGMKKRHRMIKMNPADGSMCKAETSQAEKCMMPECHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQVEKCMLPECPIDCELTEWSQWSECNKSCGKGHVIRTRMIQMEPQFGGAPCPETVQRKKCRIRKCLRNPSIQKLRWREARESRRSEQLKEESEGEQFPGCRMRPWTAWSECTKLCGGGIQERYMTVKKRFKSSQFTSCKDKKEIRACNVHPC CEA84A83F206A5A5 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 aplp1_human Amyloid beta precursor like protein 1 APLP1 Amyloid beta (A4) precursor-like protein 1 Amyloid-like protein 1 protein protein human Homo sapiens Human MGPASPAARGLSRRPGQPPLPLLLPLLLLLLRAQPAIGSLAGGSPGAAEAPGSAQVAGLCGRLTLHRDLRTGRWEPDPQRSRRCLRDPQRVLEYCRQMYPELQIARVEQATQAIPMERWCGGSRSGSCAHPHHQVVPFRCLPGEFVSEALLVPEGCRFLHQERMDQCESSTRRHQEAQEACSSQGLILHGSGMLLPCGSDRFRGVEYVCCPPPGTPDPSGTAVGDPSTRSWPPGSRVEGAEDEEEEESFPQPVDDYFVEPPQAEEEEETVPPPSSHTLAVVGKVTPTPRPTDGVDIYFGMPGEISEHEGFLRAKMDLEERRMRQINEVMREWAMADNQSKNLPKADRQALNEHFQSILQTLEEQVSGERQRLVETHATRVIALINDQRRAALEGFLAALQADPPQAERVLLALRRYLRAEQKEQRHTLRHYQHVAAVDPEKAQQMRFQVHTHLQVIEERVNQSLGLLDQNPHLAQELRPQIQELLHSEHLGPSELEAPAPGGSSEDKGGLQPPDSKDDTPMTLPKGSTEQDAASPEKEKMNPLEQYERKVNASVPRGFPFHSSEIQRDELAPAGTGVSREAVSGLLIMGAGGGSLIVLSMLLLRRKKPYGAISHGVVEVDPMLTLEEQQLRELQRHGYENPTYRFLEERP DIP protein P51693 original sequence version: 82 DIP protein P51693-1 original sequence version: 124 B95F0F4D1C5CBAC7 spon1_rat Spondin-1 Spon1 Sponf F-spondin protein protein rat Rattus norvegicus (Rat) Rat MRLSPAPLRLSRGPALLALALPLAAALAFSDETLDKVAKSEGYCSRILRAQGTRREGYTEFSLRVEGDPDFYKPGSSYRVTLSAAPPSYFRGFTLIALKENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQVFWIAPPTGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDPTLDGVTDRPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPSWQPVNVRAAPSAEFSVDRTRHLMSFLTMMGPSPDWNVGLSAEDLCTKECGWVQKVVQDLIPWDAGTDSGVTYESPNKPTIPQEKIRPLTSLDHPQSPFYDPEGGSITQVARVVIERIARKGEQCNIVPDNVDDIVADLAPEEKDEDDTPETCIYSNWSPWSACSSSTCEKGKRMRQRMLKAQLDLSVPCPDTQDFQPCMGPGCSDEDGSTCTMSEWITWSPCSVSCGMGMRSRERYVKQFPEDGSVCMLPTEETEKCTVNEECSPSSCLVTEWGEWDDCSATCGMGMKKRHRMVKMSPADGSMCKAETSQAEKCMMPECHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQAEKCMLPECPIDCELSEWSQWSECNKSCGKGHMIRTRTIQMEPQFGGAPCPETVQRKKCRARKCLRSPSIQKLRWREARESRRSEQLREESDGEQFPGCRMRPWTAWSECTKLCGGGIQERYMTVKKRFKSSQFTSCKDKKEIRACNVHPC 309525F9EAFEA89A aplp1-spon1 1 4 unspecified role unspecified role prey prey chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 5 unspecified role unspecified role bait bait c-terminal ig-tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range cappd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey physical association physical association Fig 1B Ig-fusion protein with only N-term domains fo APP does not interact with SPON1. Ca2+ required for binding interaction. However, when we tested the isolated CAPPD, immobilized as a bacterially expressed GST-fusion protein, we confirmed APP binding to F-spondin in a Ca2+-dependent manner similar to the Ig-fusion protein containing the full-length extracellular sequences of APP (Fig. 1B). spon1-aplp1-2 1 5 unspecified role unspecified role prey prey chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 4 unspecified role unspecified role bait bait ig-tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey physical association physical association Fig2b 6 deletion constructs of SPON1 Ig-fusion protein used to determine essential binding region with APP. Authors state that native brain APP also specifically retained by SPON1 but do not present any data. Authors do not specify how recombinant Ig-SPON1 construct is expressed, however Ig-APP recombinant proteins were expressed in COS cells. We found that immobilized F-spondin specifically retained recombinant and native brain APP, and that the N-terminal reelin domain and the central spondin-specific region of F-spondin were essential for binding APP, whereas the C-terminal thrombospondin repeats were not required (Fig. 2B). spon1-aplp1 1 6 unspecified role unspecified role prey prey chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 4 unspecified role unspecified role bait bait n_terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ig-tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey physical association physical association Fig2 Authors do not specify how recombinant Ig-SPON1 construct is expressed, however Ig-APP recombinant proteins were expressed in COS cells. spon1-aplp2 1 3 unspecified role unspecified role prey prey chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 4 unspecified role unspecified role bait bait required_domains required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ig-tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey physical association physical association Fig2d Authors do not specify how recombinant Ig-SPON1 construct is expressed, however Ig-APP recombinant proteins were expressed in COS cells. spon1-aplp1-1 1 5 unspecified role unspecified role prey prey binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey 4 unspecified role unspecified role bait bait ig-tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey physical association physical association Fig2c Authors do not specify how recombinant Ig-SPON1 construct is expressed, however Ig-APP recombinant proteins were expressed in COS cells. We found that immobilized F-spondin specifically retained recombinant and native brain APP, and that the N-terminal reelin domain and the central spondin-specific region of F-spondin were essential for binding APP, whereas the C-terminal thrombospondin repeats were not required (Fig. 2B). aplp1-spon1-1 2 7 unspecified role unspecified role prey prey rat-brain Rattus norvegicus brain brain Brain [cerebrum] 5 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range cappd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Fig1 We used the CAPPD-GST fusion protein immobilized on glutathione-Sepharose for affinity chromatography experiments with membrane proteins that were solubilized from rat brain with 1% Triton X-100, and used immobilized GST as a negative control (data not shown). We eluted bound proteins with high salt, and identified by MS F-spondin as a major component of the proteins bound to CAPPD.