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/ ahn-2013-5 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. bovin-brain Bos taurus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-1 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. in vitro In vitro lipase assay lipase assay predetermined predetermined participant predetermined Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. A Sphingomyelinase Assay:-[N-methyl-14C]Sphingomyelin was incubated with the N-SMase from purified bovine brain homogenate. The reaction was stopped by addition of chloroform/methanol; released radioactivity was separated into the aqueous phase and counted. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-2 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. bovin-brain Bos taurus brain brain Brain [cerebrum] chromatography chromatography technology column chromatography chromatography fingerprinting peptide massfingerprinting fingerprinting Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. Fresh bovine brain was homogenized, centrifuged, resuspended and ammonium sulfate extracts extracted. The extracts were applied to a DEAE-cellulose column, active fractions were pooled and sonicated, followed by centrifugation at 100,000× g. The resulting supernatant was applied to a butyl-Toyopearl column. The protein bound to the column was eluted with buffer containing 0.2 M (NH4)2SO4. The active fractions were applied to a DEAE-5PW HPLC column. Bound protein was eluted and applied to a Phenyl-5PW HPLC column. Bound protein was eluted and applied to a Mono S cationic exchange FPLC column. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-3 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. bovin-brain Bos taurus brain brain Brain [cerebrum] chromatography chromatography technology column chromatography chromatography western blot western blot Immuno blot Immuno blot Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. Fresh bovine brain was homogenized, centrifuged, resuspended and ammonium sulfate extracts extracted. The extracts were applied to a DEAE-cellulose column, active fractions were pooled and sonicated, followed by centrifugation at 100,000× g. The resulting supernatant was applied to a butyl-Toyopearl column. The protein bound to the column was eluted with buffer containing 0.2 M (NH4)2SO4. The active fractions were applied to a DEAE-5PW HPLC column. Bound protein was eluted and applied to a Phenyl-5PW HPLC column. Bound protein was eluted and applied to a Mono S cationic exchange FPLC column. Final purification was by Mono S (Ion Exchange) FPLC (Fast protein liquid chromatography) and Superose 12 gel filtration. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-4 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. bovin-brain Bos taurus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. Results from Fig 7B, (IP of HSP60 from rat brain synaptosomes and N-SMase2 being detected by enzyme assay) have not been included as the rat brain sample has not been purified beyond synaptosom isolation using a Percoll gradient. (see Ref. 22) Similarly in Supp fig 2A the immunoprecipitate has not been further purified. N-SMase was detected by assaying the IP supernatants for N-SMase activity. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-6 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. 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 Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-7 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. 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. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. DSP as cross-linker (dithiobis (succinimidylpropionate)) The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation ahn-2013-8 Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake. 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. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Ahn K.H., Kim S.K., Choi J.M., Jung S.Y., Won J.H., Back M.J., Fu Z., Jang J.M., Ha H.C., Kim D.K. Affinomics - Interactions curated for the Affinomics consortium. The origin of transfected N-SMase 2 (plasmid pRC/N-SMase2) has been entered as Murine; as given in Reference 18 (Kim et al., 2010) which mentions "Mouse cDNA" - also provided by Yusuf Hannun, stated to be the donor of N-SMase 2 in this publication. (The authors did not reply to a request to confirm this information.) kimdk@cau.ac.kr PLoS ONE 2013 Only protein-protein interactions imex curation f1n588_bovin SMPD2 protein protein bovin Bos taurus (Bovine) Bovine MHGTECCWLAMPARGDSWGHKHCASPGQPRRGPAPAARGGKGGRGDGRGGRRRPGLSSTLRLLLSNRRRGWRGPIGSRHQGRASEKRNAESTSRVPQSGSPLPRTSSGSPHPGSPQPPPPLGSLWDRPGSSSAAVASGACTWAPRPSGERGSRAATCTQCPGRTAARPCPPPRPSLWRPNRPAPPSVPAMKPNFTLRLRVFNLNCWGIPYLSKHRADRVKRLGDFLNMESFDLALLEEVWSEQDFQYLRQKLLPTYPAAHYFRSGIIGSGLCVFSKHPIQEFTQHVFTLNGYPYMIHHCDWFCGKAVGLLVLHLSGLVLNAYVTHLHAEYNRQKDTYLAHRVAQAWELAQFIHHTSKKADVVLLCGDLNLHPKDLGCRLLKEWTGLHDAYLETRDFKGSEEGCTMVPKNCYVKHQELGPFPLGIRIDYVLYKAVSGLYIFCKTFKTTTGHDPYSGPPFSDHEALMATLCVRHSPPQHNPSPTHGPAESSPLTSVLREAWAEVDQGMAQAHWWATMAGYAVGLGLLLLALLCALAAGGWIREAALLLWTPSVGLVLGAGAFYLFHVQEAKGLCRTRAELQHVLGRAREAQDLGPESQPALLLGQQERERAEEQ ch60_bovin 60 kDa heat shock protein, mitochondrial HSPD1 HSP60 60 kDa chaperonin Chaperonin 60 Heat shock protein 60 Mitochondrial matrix protein P1 protein protein bovin Bos taurus (Bovine) Bovine MLRLPAVLRQMRPVSRALALHLTRAYAKDVKFGADARALMLQGVDLLADAVAVTMGPKGRTVIIEQSWGSPRVTKDGVTVAKSIDLKDKYKNIGAKLVQDVANNTNEEAGDGTTTATVLARSIAKEGFEKISKGANPVEIRRGVMLAVDAVIVELKKQSKPVTTPEEIAQVATISANGDKEIGNIISDAMKKVGRKGVITVKDGKTLNDELEIIEGMKFDRGYISPYFINTSKGQKCEFQDAYVLLSEKKISSVQSIVPALEIANAHRKPLVIIAEDVDGEALSTLVLNRLKVGLQVVAVKAPGFGDNRKNQLKDMAIATGGAVFGEEGLNLNLEDVQPHDLGKVGEVIVTKDDAMLLKGKGDKAQIEKRIQEIIEQLDITTSEYEKEKLNERLAKLSDGVAVLKVGGTSDVEVNEKKDRVTDALNATRAAVEEGIVLGGDCALLRCIPALESITPANEDQKTGIEIIKKTLKIPAMTIAKNAGVEGSLIVEKIMQSSSEVGYDAMLGDFVNMVEKGIIDPTKVVRTALLDAAGVASLLTTAEVVVTEIPKEEKDPGMGGMGGMGGGMGGGMF sphingomyelin sphingomyelin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ch60_human 60 kDa heat shock protein, mitochondrial HSPD1 HSP60 Heat shock protein 60 60 kDa chaperonin Chaperonin 60 Mitochondrial matrix protein P1 P60 lymphocyte protein HuCHA60 protein protein human Homo sapiens Human MLRLPTVFRQMRPVSRVLAPHLTRAYAKDVKFGADARALMLQGVDLLADAVAVTMGPKGRTVIIEQSWGSPKVTKDGVTVAKSIDLKDKYKNIGAKLVQDVANNTNEEAGDGTTTATVLARSIAKEGFEKISKGANPVEIRRGVMLAVDAVIAELKKQSKPVTTPEEIAQVATISANGDKEIGNIISDAMKKVGRKGVITVKDGKTLNDELEIIEGMKFDRGYISPYFINTSKGQKCEFQDAYVLLSEKKISSIQSIVPALEIANAHRKPLVIIAEDVDGEALSTLVLNRLKVGLQVVAVKAPGFGDNRKNQLKDMAIATGGAVFGEEGLTLNLEDVQPHDLGKVGEVIVTKDDAMLLKGKGDKAQIEKRIQEIIEQLDVTTSEYEKEKLNERLAKLSDGVAVLKVGGTSDVEVNEKKDRVTDALNATRAAVEEGIVLGGGCALLRCIPALDSLTPANEDQKIGIEIIKRTLKIPAMTIAKNAGVEGSLIVEKIMQSSSEVGYDAMAGDFVNMVEKGIIDPTKVVRTALLDAAGVASLLTTAEVVVTEIPKEEKDPGMGAMGGMGGGMGGGMF nsma2_mouse Sphingomyelin phosphodiesterase 3 Smpd3 Neutral sphingomyelinase 2 Neutral sphingomyelinase II protein protein mouse Mus musculus Mouse MVLYTTPFPNSCLSALHAVSWALIFPCYWLVDRLLASFIPTTYEKRQRADDPCCLQLFCTVLFTPVYLALLVAALPFAFLGFIFWSPLQSARRPYSYSRLEDKNPAGGAALLSEWKGTGAGKSFCFATANVCLLPDSLARLNNVFNTQARAKEIGQRIRNGAARPQIKIYIDSPTNTSISAASFSSLVSPQGGDGSRAVPGSIKRTASVEYKGDGGRHPSDEAANGPASGEQADGSLEDSCIVRIGGEEGGRPQEADDPAAGSQARNGAGGTPKGQTPNHNQRDGDSGSLGSPSASRESLVKARAGQDSGGSGEPGANSKLLYKTSVVKKAAARRRRHPDEAFDHEVSAFFPANLDFLCLQEVFDKRAAAKLKEQLHGYFEYILYDVGVYGCHGCCNFKCLNSGLFFASRYPVMDVAYHCYPNGCSFDALASKGALFLKVQVGSTPQDQRIVGYIACTHLHAPPEDSAVRCEQLDLLQDWLADFRKSTSSTSTANPEELVVFDVICGDLNFDNCSSDDKLEQQHSLFTRYKDPCRLGPGEEKPWAIGTLLDTNGLYDEDVCTPDNLQKVLESEEGRREYLAFPTSKSPGAGQKGRKDLLKGNGRRIDYMLHAEEGLCPDWKAEVEEFSFITQLSGLTDHLPVAMRLMVSAGEEEA hspd1-smpd2 1567340 n/a 1567348 unspecified role unspecified role prey prey n/a 1567349 unspecified role unspecified role bait bait physical association physical association false false false Figure 4C smpd2-hspd1 1567340 n/a 1567348 unspecified role unspecified role bait bait n/a 1567349 unspecified role unspecified role prey prey physical association physical association false false false Figure 4C smpd2-sphingomyelin 1567341 n/a 1567348 enzyme enzyme neutral component neutral component bovin-brain Bos taurus brain brain Brain [cerebrum] n/a 1567350 enzyme target enzyme target substrate substrate neutral component neutral component n-methyl_choline 14c radiolabel 14c radiolabel C14 14C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false lipid cleavage lipid cleavage false false false Figure 1,2,3,5 Ca 2+, Mg2+ and Mn2+ phosphatidylserine GW4869 (a cell-permeable, non-competitive inhibitor of neutral sphingomyelinases) smpd2-sphingomyelin-1 1567342 n/a 1567348 unspecified role unspecified role neutral component neutral component n/a 1567349 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 1D smpd2-sphingomyelin-2 1567343 n/a 1567349 unspecified role unspecified role neutral component neutral component n/a 1567348 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 3 hspd1-smpd2-1 1567344 n/a 1567348 unspecified role unspecified role prey prey n/a 1567349 unspecified role unspecified role bait bait physical association physical association false false false Figure 4A,5 hspd1-smpd2-2 1567345 n/a 1567351 unspecified role unspecified role bait bait n/a 1567352 unspecified role unspecified role prey prey physical association physical association false false false Figure 6 hspd1-smpd2-3 1567346 n/a 1567352 unspecified role unspecified role neutral component neutral component n/a 1567351 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 6 hspd1-smpd3 1567347 n/a 1567351 unspecified role unspecified role neutral component neutral component n/a 1567352 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 8B, supp 3.