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/ matsuda-2013-1 Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. 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 Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation matsuda-2013-2 Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. 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 Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation matsuda-2013-3 Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. 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 Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation matsuda-2013-4 Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. 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 Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation matsuda-2013-5 Prolyl oligopeptidase is a glyceraldehyde-3-phosphate dehydrogenase-binding protein that regulates genotoxic stress-induced cell death. 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 Int. J. Biochem. Cell Biol. (1357-2725) 2013 Only protein-protein interactions imex curation 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 ppce_human Prolyl endopeptidase PREP PEP Post-proline cleaving enzyme protein protein human Homo sapiens Human MLSLQYPDVYRDETAVQDYHGHKICDPYAWLEDPDSEQTKAFVEAQNKITVPFLEQCPIRGLYKERMTELYDYPKYSCHFKKGKRYFYFYNTGLQNQRVLYVQDSLEGEARVFLDPNILSDDGTVALRGYAFSEDGEYFAYGLSASGSDWVTIKFMKVDGAKELPDVLERVKFSCMAWTHDGKGMFYNSYPQQDGKSDGTETSTNLHQKLYYHVLGTDQSEDILCAEFPDEPKWMGGAELSDDGRYVLLSIREGCDPVNRLWYCDLQQESSGIAGILKWVKLIDNFEGEYDYVTNEGTVFTFKTNRQSPNYRVINIDFRDPEESKWKVLVPEHEKDVLEWIACVRSNFLVLCYLHDVKNILQLHDLTTGALLKTFPLDVGSIVGYSGQKKDTEIFYQFTSFLSPGIIYHCDLTKEELEPRVFREVTVKGIDASDYQTVQIFYPSKDGTKIPMFIVHKKGIKLDGSHPAFLYGYGGFNISITPNYSVSRLIFVRHMGGILAVANIRGGGEYGETWHKGGILANKQNCFDDFQCAAEYLIKEGYTSPKRLTINGGSNGGLLVAACANQRPDLFGCVIAQVGVMDMLKFHKYTIGHAWTTDYGCSDSKQHFEWLVKYSPLHNVKLPEADDIQYPSMLLLTADHDDRVVPLHSLKFIATLQYIVGRSRKQSNPLLIHVDTKAGHGAGKPTAKVIEEVSDMFAFIARCLNVDWIP prep-gapdh 1546612 n/a 1546617 unspecified role unspecified role prey prey human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC n/a 1546618 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1B prep-gapdh-1 1546613 n/a 1546618 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 false ecoli Escherichia coli (strain K12) n/a 1546617 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 false false false 1A gapdh-prep 1546614 n/a 1546618 unspecified role unspecified role neutral component neutral component n/a 1546617 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 2A prep-gapdh-2 1546615 n/a 1546618 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 false n/a 1546617 unspecified role unspecified role prey prey physical association physical association false false false 2B prep-gapdh-3 1546616 n/a 1546617 unspecified role unspecified role prey prey n/a 1546618 unspecified role unspecified role bait bait proximity proximity close-contact colocalization neighbourhood interaction false false false 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.