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/ Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Int. J. Biochem. Cell Biol. (1357-2725) 2013 imex curation Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions human-nb1 Human NB-1 Neuroblastoma cell line nb1 NB-1 Neuroblastoma cell line IFO50295 - JCRB Cell Bank anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp Accepted 2014-JUN-25 by ORCHARD Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Int. J. Biochem. Cell Biol. (1357-2725) 2013 imex curation Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions human-nb1 Human NB-1 Neuroblastoma cell line nb1 NB-1 Neuroblastoma cell line IFO50295 - JCRB Cell Bank pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp Accepted 2014-JUN-25 by ORCHARD Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Int. J. Biochem. Cell Biol. (1357-2725) 2013 imex curation Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down partial id prot seq partial identification of protein sequence partial id prot seq Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. Purified rhPOP was coupled to an NHS-activated HiTrap column and NB-1 cell extract loaded. Bound proteins were eluted with NaCl and analysed by SDS-PAGE, blotted onto PVDF membranes and stained bands analysed with a protein sequencer. It is stated in the Methods section that the GST tag has been removed from the POP fusion protein by thrombin cleavage. However according to the Novagen web site for the pET-41 Ek/LIC vector the thrombin cleavage site is downstream of a His tag, so an S-tag is retained as an N-terminal tag. sakaguti@gly.oups.ac.jp Accepted 2014-JUN-25 by ORCHARD Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Int. J. Biochem. Cell Biol. (1357-2725) 2013 imex curation Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions human-nb1 Human NB-1 Neuroblastoma cell line nb1 NB-1 Neuroblastoma cell line IFO50295 - JCRB Cell Bank confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp Accepted 2014-JUN-25 by ORCHARD Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. Int. J. Biochem. Cell Biol. (1357-2725) 2013 imex curation Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. sakaguti@gly.oups.ac.jp 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Matsuda T., Sakaguchi M., Tanaka S., Yoshimoto T., Takaoka M. Affinomics - Interactions curated for the Affinomics consortium. It is stated in the Methods section that the GST tag has been removed from the POP fusion protein by thrombin cleavage. However according to the Novagen web site for the pET-41 Ek/LIC vector the thrombin cleavage site is downstream of a His tag, so an S-tag is retained as an N-terminal tag. sakaguti@gly.oups.ac.jp Accepted 2014-JUN-25 by ORCHARD Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation ppce_human Prolyl endopeptidase PREP PEP Post-proline cleaving enzyme protein protein human Homo sapiens Human MLSLQYPDVYRDETAVQDYHGHKICDPYAWLEDPDSEQTKAFVEAQNKITVPFLEQCPIRGLYKERMTELYDYPKYSCHFKKGKRYFYFYNTGLQNQRVLYVQDSLEGEARVFLDPNILSDDGTVALRGYAFSEDGEYFAYGLSASGSDWVTIKFMKVDGAKELPDVLERVKFSCMAWTHDGKGMFYNSYPQQDGKSDGTETSTNLHQKLYYHVLGTDQSEDILCAEFPDEPKWMGGAELSDDGRYVLLSIREGCDPVNRLWYCDLQQESSGIAGILKWVKLIDNFEGEYDYVTNEGTVFTFKTNRQSPNYRVINIDFRDPEESKWKVLVPEHEKDVLEWIACVRSNFLVLCYLHDVKNILQLHDLTTGALLKTFPLDVGSIVGYSGQKKDTEIFYQFTSFLSPGIIYHCDLTKEELEPRVFREVTVKGIDASDYQTVQIFYPSKDGTKIPMFIVHKKGIKLDGSHPAFLYGYGGFNISITPNYSVSRLIFVRHMGGILAVANIRGGGEYGETWHKGGILANKQNCFDDFQCAAEYLIKEGYTSPKRLTINGGSNGGLLVAACANQRPDLFGCVIAQVGVMDMLKFHKYTIGHAWTTDYGCSDSKQHFEWLVKYSPLHNVKLPEADDIQYPSMLLLTADHDDRVVPLHSLKFIATLQYIVGRSRKQSNPLLIHVDTKAGHGAGKPTAKVIEEVSDMFAFIARCLNVDWIP 139072B990820B90 g3p_human Glyceraldehyde-3-phosphate dehydrogenase GAPDH OK/SW-cl.12 CDABP0047 GAPD Peptidyl-cysteine S-nitrosylase GAPDH protein protein human Homo sapiens Human MGKVKVGVNGFGRIGRLVTRAAFNSGKVDIVAINDPFIDLNYMVYMFQYDSTHGKFHGTVKAENGKLVINGNPITIFQERDPSKIKWGDAGAEYVVESTGVFTTMEKAGAHLQGGAKRVIISAPSADAPMFVMGVNHEKYDNSLKIISNASCTTNCLAPLAKVIHDNFGIVEGLMTTVHAITATQKTVDGPSGKLWRDGRGALQNIIPASTGAAKAVGKVIPELNGKLTGMAFRVPTANVSVVDLTCRLEKPAKYDDIKKVVKQASEGPLKGILGYTEHQVVSSDFNSDTHSSTFDAGAGIALNDHFVKLISWYDNEFGYSNRVVDLMAHMASKE DIP protein P04406 original sequence version: null DIP protein P04406 original sequence version: 196 DIP protein P04406 original sequence version: 198 DIP protein P04406 original sequence version: 156 C9C135E8AE3E8744 prep-gapdh 5 7 unspecified role unspecified role prey prey human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC 6 unspecified role unspecified role bait bait n_terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction 1B prep-gapdh-1 3 6 unspecified role unspecified role bait bait n_terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey human-nb1 Human NB-1 Neuroblastoma cell line nb1 NB-1 Neuroblastoma cell line IFO50295 - JCRB Cell Bank physical association physical association 1A gapdh-prep 4 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization 2A prep-gapdh-2 1 6 unspecified role unspecified role bait bait n_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role prey prey physical association physical association 2B prep-gapdh-3 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait proximity proximity close-contact colocalization neighbourhood interaction Figure 6 An interaction in the cytoplasm was detected only when cells were treated with both Ara-C and SUAM-14746, inhibiting GAPDH nuclear translocation. SUAM-14746 - a POP inhibitor. Ara-C (cytosine arabinoside) -a pyrimidine nucleoside analog that inhibits the synthesis of DNA. It induces cell cycle arrest in the S phase and can induce apoptosis.