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/ shu-2007a-3 GRP78 and Raf-1 cooperatively confer resistance to endoplasmic reticulum stress-induced apoptosis. human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 cosedimentation cosedimentation western blot western blot Immuno blot Immuno blot Shu CW., Sun FC., Cho JH., Lin CC., Liu PF., Chen PY., Chang MD., Fu HW., Lai YK. Apoptosis - Interactions involving proteins with a function related to apoptosis yklai@life.nthu.edu.tw J. Cell. Physiol. (0021-9541) 2007 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer shu-2007a-2 GRP78 and Raf-1 cooperatively confer resistance to endoplasmic reticulum stress-induced apoptosis. human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Shu CW., Sun FC., Cho JH., Lin CC., Liu PF., Chen PY., Chang MD., Fu HW., Lai YK. Apoptosis - Interactions involving proteins with a function related to apoptosis yklai@life.nthu.edu.tw J. Cell. Physiol. (0021-9541) 2007 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer shu-2007a-1 GRP78 and Raf-1 cooperatively confer resistance to endoplasmic reticulum stress-induced apoptosis. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Shu CW., Sun FC., Cho JH., Lin CC., Liu PF., Chen PY., Chang MD., Fu HW., Lai YK. Apoptosis - Interactions involving proteins with a function related to apoptosis yklai@life.nthu.edu.tw J. Cell. Physiol. (0021-9541) 2007 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer bip_human Endoplasmic reticulum chaperone BiP HSPA5 GRP78 Immunoglobulin heavy chain-binding protein 78 kDa glucose-regulated protein Binding-immunoglobulin protein Heat shock protein 70 family protein 5 Heat shock protein family A member 5 protein protein human Homo sapiens Human MKLSLVAAMLLLLSAARAEEEDKKEDVGTVVGIDLGTTYSCVGVFKNGRVEIIANDQGNRITPSYVAFTPEGERLIGDAAKNQLTSNPENTVFDAKRLIGRTWNDPSVQQDIKFLPFKVVEKKTKPYIQVDIGGGQTKTFAPEEISAMVLTKMKETAEAYLGKKVTHAVVTVPAYFNDAQRQATKDAGTIAGLNVMRIINEPTAAAIAYGLDKREGEKNILVFDLGGGTFDVSLLTIDNGVFEVVATNGDTHLGGEDFDQRVMEHFIKLYKKKTGKDVRKDNRAVQKLRREVEKAKRALSSQHQARIEIESFYEGEDFSETLTRAKFEELNMDLFRSTMKPVQKVLEDSDLKKSDIDEIVLVGGSTRIPKIQQLVKEFFNGKEPSRGINPDEAVAYGAAVQAGVLSGDQDTGDLVLLDVCPLTLGIETVGGVMTKLIPRNTVVPTKKSQIFSTASDNQPTVTIKVYEGERPLTKDNHLLGTFDLTGIPPAPRGVPQIEVTFEIDVNGILRVTAEDKGTGNKNKITITNDQNRLTPEEIERMVNDAEKFAEEDKKLKERIDTRNELESYAYSLKNQIGDKEKLGGKLSSEDKETMEKAVEEKIEWLESHQDADIEDFKAKKKELEEIVQPIISKLYGSAGPPPTGEEDTAEKDEL 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 raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF pten_human Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Phosphatase and tensin homolog Mutated in multiple advanced cancers 1 PTEN MMAC1 TEP1 protein protein human Homo sapiens Human MTAIIKEIVSRNKRRYQEDGFDLDLTYIYPNIIAMGFPAERLEGVYRNNIDDVVRFLDSKHKNHYKIYNLCAERHYDTAKFNCRVAQYPFEDHNPPQLELIKPFCEDLDQWLSEDDNHVAAIHCKAGKGRTGVMICAYLLHRGKFLKAQEALDFYGEVRTRDKKGVTIPSQRRYVYYYSYLLKNHLDYRPVALLFHKMMFETIPMFSGGTCNPQFVVCQLKVKIYSSNSGPTRREDKFMYFEFPQPLPVCGDIKVEFFHKQNKMLKKDKMFHFWVNTFFIPGPEETSEKVENGSLCDQEIDSICSIERADNDKEYLVLTLTKNDLDKANKDKANRYFSPNFKVKLYFTKTVEEPSNPEASSSTSVTPDVSDNEPDHYRYSDTTDSDPENEPFDEDQHTQITKV hspa5-raf1-2 Raf-1, GRP78 and HSP60 colocalize in mitochondrial fraction of H460 cells 693847 N/A 693850 unspecified role unspecified role neutral component neutral component N/A 693851 unspecified role unspecified role neutral component neutral component N/A 693852 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 5 D Phosphorylation of Raf-1 at Ser338 and/or Tyr340 in mitochondria, reported to block apoptosis, was markedly diminished in AEBSF-pretreated cells during ER stress (caused by TG treatment). hspa5-pten GRP78 and PTEN colocalize in cytoplasmic fraction of H460 cells 693847 N/A 693850 unspecified role unspecified role neutral component neutral component N/A 693853 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 5 D hspa5-raf1-3 GRP78 and HSP60 colocalize in microsomal fraction of H460 cells 693847 N/A 693852 unspecified role unspecified role neutral component neutral component N/A 693850 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 5 D The microsomal fraction contains the ER fraction. hspa5-raf1-1 GRP78 coimmunoprecipitates RAF-1 H460 lysates 693848 N/A 693850 unspecified role unspecified role bait bait N/A 693852 unspecified role unspecified role prey prey physical association physical association false false false Fig. 5 C raf1-hspa5 GRP78 coimmunoprecipitates with RAF-1 H460 lysates 693848 N/A 693852 unspecified role unspecified role bait bait N/A 693850 unspecified role unspecified role prey prey physical association physical association false false false Fig. 5 C hspa5-raf1 His-GRP78 pulls down RAF-1 from H460 extracts 693849 N/A 693852 unspecified role unspecified role prey prey human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 N/A 693850 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false Fig. 5 B GRP78 interacted with Raf-1 in both untreated and TG-treated cells. In the presence of ATP, GRP78 bound to Raf-1 with a lower affinity, implying that ATP stimulated Raf-1 release from GRP78.