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/ TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. p.hyslop@utoronto.ca Accepted 2006-SEP-13 by ORCHARD. Nature (0028-0836) 2006 TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. 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 anti tag western anti tag western blot anti tag western Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. p.hyslop@utoronto.ca Accepted 2006-SEP-13 by ORCHARD. Nature (0028-0836) 2006 TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. 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. bn-page blue native page bn-page predetermined predetermined participant predetermined Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. The proteins in CHAPSO-solubilized membrane fractions were separated on a 5-13% Blue Native polyacrylamide gel, followed by a second dimension on a NuPAGE BisTris 4�12% precast 2-D gel (Invitrogen) for SDS-PAGE. p.hyslop@utoronto.ca Accepted 2006-SEP-13 by ORCHARD. Nature (0028-0836) 2006 TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. 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 Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca Accepted 2006-SEP-14 by ORCHARD. Nature (0028-0836) 2006 Only protein-protein interactions imex curation TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. 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. cosedimentation cosedimentation western blot western blot Immuno blot Immuno blot Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. 1 ml of total protein extracts was applied to the top of an 11.0-ml 10�40% (w/v) linear glycerol gradient containing 25 mM HEPES, pH 7.2, 150 mM NaCl, and 0.5% CHAPSO. Gradients were centrifuged for 15 hours at 35,000 rpm and 4 �C using a Beckman SW41 rotor and were collected into 1.0-ml fractions from the top of the centrifugation tube. p.hyslop@utoronto.ca Accepted 2006-SEP-13 by ORCHARD. Nature (0028-0836) 2006 TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-blastocyst Mus musculus blastocyst blastocyst blastocyst anti bait coip anti bait coimmunoprecipitation anti bait coip ms participant Identification by mass spectrometry ms participant Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca Accepted 2006-SEP-14 by ORCHARD. 24-JAN-2007: Contacted by JYOTI. Nature (0028-0836) 2006 Only protein-protein interactions imex curation TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature (0028-0836) 2006 imex curation Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. Synapse - Interactions of proteins with an established role in the presynapse. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology the information in paras 1 and 2 is found in chen-2006a-7 PS1 complexes were immunoprecipitated from CHAPSO-solubilised membranes of mouse blastocyst-derived cells and components were identified by mass spectrometry. p.hyslop@utoronto.ca 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen F., Hasegawa H., Schmitt-Ulms G., Kawarai T., Bohm C., Katayama T., Gu Y., Sanjo N., Glista M., Rogaeva E., Wakutani Y., Pardossi-Piquard R., Ruan X., Tandon A., Checler F., Marambaud P., Hansen K., Westaway D., St George-Hyslop P., Fraser P. p.hyslop@utoronto.ca Accepted 2006-SEP-13 by ORCHARD. Nature (0028-0836) 2006 tmeda_human Transmembrane emp24 domain-containing protein 10 21 kDa transmembrane-trafficking protein Transmembrane protein Tmp21 Tmp-21-I p24delta S31III125 TMED10 TMP21 p23 p24 family protein delta-1 S31I125 protein protein human Homo sapiens Human MSGLSGPPARRGPFPLALLLLFLLGPRLVLAISFHLPINSRKCLREEIHKDLLVTGAYEISDQSGGAGGLRSHLKITDSAGHILYSKEDATKGKFAFTTEDYDMFEVCFESKGTGRIPDQLVILDMKHGVEAKNYEEIAKVEKLKPLEVELRRLEDLSESIVNDFAYMKKREEEMRDTNESTNTRVLYFSIFSMFCLIGLATWQVFYLRRFFKAKKLIE 0A0486BE65C4DBBB psn1_mouse Presenilin-1 Protein S182 Psen1 Ad3h Psnl1 protein protein mouse Mus musculus Mouse MTEIPAPLSYFQNAQMSEDSHSSSAIRSQNDSQERQQQHDRQRLDNPEPISNGRPQSNSRQVVEQDEEEDEELTLKYGAKHVIMLFVPVTLCMVVVVATIKSVSFYTRKDGQLIYTPFTEDTETVGQRALHSILNAAIMISVIVIMTILLVVLYKYRCYKVIHAWLIISSLLLLFFFSFIYLGEVFKTYNVAVDYVTVALLIWNFGVVGMIAIHWKGPLRLQQAYLIMISALMALVFIKYLPEWTAWLILAVISVYDLVAVLCPKGPLRMLVETAQERNETLFPALIYSSTMVWLVNMAEGDPEAQRRVPKNPKYNTQRAERETQDSGSGNDDGGFSEEWEAQRDSHLGPHRSTPESRAAVQELSGSILTSEDPEERGVKLGLGDFIFYSVLVGKASATASGDWNTTIACFVAILIGLCLTLLLLAIFKKALPALPISITFGLVFYFATDYLVQPFMDQLAFHQFYI DIP protein P49769 original sequence version: 150 DIP protein P49769 original sequence version: 110 D07215B4BAD2D549 tmed2_human Transmembrane emp24 domain-containing protein 2 TMED2 RNP24 Membrane protein p24A p24 family protein beta-1 p24 protein protein human Homo sapiens Human MVTLAELLVLLAALLATVSGYFVSIDAHAEECFFERVTSGTKMGLIFEVAEGGFLDIDVEITGPDNKGIYKGDRESSGKYTFAAHMDGTYKFCFSNRMSTMTPKIVMFTIDIGEAPKGQDMETEAHQNKLEEMINELAVAMTAVKHEQEYMEVRERIHRAINDNTNSRVVLWSFFEALVLVAMTLGQIYYLKRFFEVRRVV C452370E459DC894 tmed2_mouse Transmembrane emp24 domain-containing protein 2 Sid 394 Tmed2 Rnp24 Sid394 Membrane protein p24A p24 family protein beta-1 COPI-coated vesicle membrane protein p24 protein protein mouse Mus musculus Mouse MVTLAELLALLAALLATASGYFVSIDAHAEECFFERVTSGTKMGLIFEVAEGGFLDIDVEITGPDNKGIYKGDRESSGKYTFAAHMDGTYKFCFSNRMSTMTPKIVMFTIDIGEAPKGQDMETEAHQNKLEEMINELAVAMTAVKHEQEYMEVRERIHRAINDNTNSRVVLWSFFEALVLVAMTLGQIYYLKRFFEVRRVV F5DE259FC419CF85 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 pen2_human Gamma-secretase subunit PEN-2 Presenilin enhancer protein 2 PSENEN PEN2 MDS033 protein protein human Homo sapiens Human MNLERVSNEEKLNLCRKYYLGGFAFLPFLWLVNIFWFFREAFLVPAYTEQSQIKGYVWRSAVGFLFWVIVLTSWITIFQIYRPRWGALGDYLSFTIPLGTP DIP protein Q9NZ42 original sequence version: 125 FDBA4D299488EA02 nica_mouse Nicastrin Ncstn protein protein mouse Mus musculus Mouse MATTRGGSGPDPGSRGLLLLSFSVVLAGLCGGNSVERKIYIPLNKTAPCVRLLNATHQIGCQSSISGDTGVIHVVEKEEDLKWVLTDGPNPPYMVLLEGKLFTRDVMEKLKGTTSRIAGLAVTLAKPNSTSSFSPSVQCPNDGFGIYSNSYGPEFAHCKKTLWNELGNGLAYEDFSFPIFLLEDENETKVIKQCYQDHNLGQNGSAPSFPLCAMQLFSHMHAVISTATCMRRSFIQSTFSINPEIVCDPLSDYNVWSMLKPINTSVGLEPDVRVVVAATRLDSRSFFWNVAPGAESAVASFVTQLAAAEALHKAPDVTTLSRNVMFVFFQGETFDYIGSSRMVYDMENGKFPVRLENIDSFVELGQVALRTSLDLWMHTDPMSQKNESVKNQVEDLLATLEKSGAGVPEVVLRRLAQSQALPPSSLQRFLRARNISGVVLADHSGSFHNRYYQSIYDTAENINVTYPEWQSPEEDLNFVTDTAKALANVATVLARALYELAGGTNFSSSIQADPQTVTRLLYGFLVRANNSWFQSILKHDLRSYLDDRPLQHYIAVSSPTNTTYVVQYALANLTGKATNLTREQCQDPSKVPNESKDLYEYSWVQGPWNSNRTERLPQCVRSTVRLARALSPAFELSQWSSTEYSTWAESRWKDIQARIFLIASKELEFITLIVGFSTLVFSLIVTYCINAKADVLFVAPREPGAVSY DIP protein P57716 original sequence version: 105 DIP protein P57716 original sequence version: 70 32B57BB478FEB0AC pen2_mouse Gamma-secretase subunit PEN-2 Presenilin enhancer protein 2 Psenen Pen2 protein protein mouse Mus musculus Mouse MNLERVSNEEKLNLCRKYYLGGFAFLPFLWLVNIFWFFREAFLAPAYTEQSQIKGYVWRSAVGFLFWVIILATWITIFQIYRPRWGALGDYLSFTIPLGTP DIP protein Q9CQR7 original sequence version: 102 DIP protein Q9CQR7 original sequence version: 66 6596CA2984FBCC59 nica_human Nicastrin NCSTN KIAA0253 UNQ1874/PRO4317 protein protein human Homo sapiens Human MATAGGGSGADPGSRGLLRLLSFCVLLAGLCRGNSVERKIYIPLNKTAPCVRLLNATHQIGCQSSISGDTGVIHVVEKEEDLQWVLTDGPNPPYMVLLESKHFTRDLMEKLKGRTSRIAGLAVSLTKPSPASGFSPSVQCPNDGFGVYSNSYGPEFAHCREIQWNSLGNGLAYEDFSFPIFLLEDENETKVIKQCYQDHNLSQNGSAPTFPLCAMQLFSHMHAVISTATCMRRSSIQSTFSINPEIVCDPLSDYNVWSMLKPINTTGTLKPDDRVVVAATRLDSRSFFWNVAPGAESAVASFVTQLAAAEALQKAPDVTTLPRNVMFVFFQGETFDYIGSSRMVYDMEKGKFPVQLENVDSFVELGQVALRTSLELWMHTDPVSQKNESVRNQVEDLLATLEKSGAGVPAVILRRPNQSQPLPPSSLQRFLRARNISGVVLADHSGAFHNKYYQSIYDTAENINVSYPEWLSPEEDLNFVTDTAKALADVATVLGRALYELAGGTNFSDTVQADPQTVTRLLYGFLIKANNSWFQSILRQDLRSYLGDGPLQHYIAVSSPTNTTYVVQYALANLTGTVVNLTREQCQDPSKVPSENKDLYEYSWVQGPLHSNETDRLPRCVRSTARLARALSPAFELSQWSSTEYSTWTESRWKDIRARIFLIASKELELITLTVGFGILIFSLIVTYCINAKADVLFIAPREPGAVSY DIP protein Q92542 original sequence version: 74 DIP protein Q92542 original sequence version: 146 C8C0EAEAD89E976A tmed10-psen1 4 8 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey association association 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 TMP21 also interacts with the p24 cargo protein p24a, but as a component of a separate complex. TMP21 also coprecipitates with a APH-1 but the authors do not distinguish between APH-1 alpha (Q96BI3) and APH-1 beta (Q8WW43). TMP21 coprecipitates with p24a in this experimet but whereas the other proteins coprecipitated are all part of the same complex, p24a is not. Note there are separate bands for mature nicastrin and immature nicastrin. It is not clear but it seems these interactions take place in HEK-293 cells. tmed10-tmed2 4 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 1 B p24a, another member of the p24 cargo protein family, does not interact with any members of the presenilin complex but does co-precipitate TMP21. It is not clear but it seems this interaction is in HEK-293 cells. tmed10-psen1-1 7 12 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed10-ncstn 7 8 unspecified role unspecified role prey prey 16 unspecified role unspecified role bait bait physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed10-psenen 7 13 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed2-psen1 1 9 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed2-ncstn 1 11 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed2-psenen 1 11 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 tmed10-psen1-3 2 12 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 It is not clear what the species of the flag-tagged TMP21 is, but it is assumed it is human. tmed10-ncstn-2 2 16 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 It is not clear what the species of the flag-tagged TMP21 is, but it is assumed it is human. tmed10-psenen-2 2 13 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Supp. Fig. 1 A endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 It is not clear what the species of the flag-tagged TMP21 is, but it is assumed it is human. tmed10-psen1-4 5 16 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component physical association physical association Supp. Fig. 1 B Note APH-1 also cosediments with these proteins but the authors do not distighish between APH-1 alpha and APH-1 beta. endogenous TMP21 from mouse brain, neuron-like SHSY-5Y cells and human embryonic kidney (HEK-293) cells could be co-immunoprecipitated with endogenous nicastrin, aph-1, pen-2 and PS1 (Fig. 1a, and Supplementary Fig. 1a) and that it had an overlapping size distribution with presenilin complex components in high-molecular-mass (more than 650-kDa) fractions on glycerol velocity gradients tmed10-psen1-5 3 10 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 16 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component physical association physical association Supp. Fig. 1 C Note APH-1 also comigrates with these proteins but the authors do not distighish between APH-1 alpha and APH-1 beta. On 2D-gel electrophoresis in wild type cells expressing endogenous proteins, TMP21 is distributed into a series of peaks, some of which overlap those of the mature, functional ~660 kDa PS1 complex, the ~440 kDa immature non-functional complex, and the ~150 kDa nicastrin-aph-1 complex. About 25% of the TMP21 signal resides in the ~660 kDa complex. In contrast, in both PS1/PS2 double knockout cells and in pen-2 knock-down cells, TMP21 is destabilized from the ~660 kDa complex (<5% of signal intensity), and instead, TMP21 is predominantly localized with the ~150 kDa nicastrin:aph-1 complex and in a 30 kDa complex. In both PS1/PS2 double knockout cells and in pen-2 knock-down cells, TMP21 is destabilized from the ~660 kDa complex (<5% of signal intensity), and instead, TMP21 is predominantly localized with the ~150 kDa nicastrin:aph-1 complex and in a 30 kDa complex. psen1-ncstn 6 9 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey association association Note APH-1 also cosediments with these proteins but the authors do not distighish between APH-1 alpha and APH-1 beta. All four known constituents of the PS1 complex (nicastrin3, aph-1 (refs 4, 5), pen-2 (ref. 5) and PS1 (ref. 1)) were present only in the co-precipitates from wild-type cells. The complex was found in wild-type cells and not in PS1-PS2 double knockout cells.