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/ OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. Mol. Cell (1097-2765) 2005 imex curation Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer gsemenza@jhmi.edu 23-JAN-2007: Contacted by JYOTI. A human brain cDNA library was used. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis gsemenza@jhmi.edu Accepted 2006-Not a month!-18 by DAVET. 23-JAN-2007: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell (1097-2765) 2005 Only protein-protein interactions imex curation OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. Mol. Cell (1097-2765) 2005 imex curation Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer gsemenza@jhmi.edu 23-JAN-2007: Contacted by JYOTI. A human brain cDNA library was used. 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis gsemenza@jhmi.edu Accepted 2006-Not a month!-18 by DAVET. 23-JAN-2007: Contacted by JYOTI. Mol. Cell (1097-2765) 2005 Only protein-protein interactions imex curation OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. Mol. Cell (1097-2765) 2005 imex curation Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer gsemenza@jhmi.edu 23-JAN-2007: Contacted by JYOTI. A human brain cDNA library was used. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis gsemenza@jhmi.edu Accepted 2006-Not a month!-18 by DAVET. 23-JAN-2007: Contacted by JYOTI. Mol. Cell (1097-2765) 2005 Only protein-protein interactions imex curation OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. Mol. Cell (1097-2765) 2005 imex curation Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer gsemenza@jhmi.edu 23-JAN-2007: Contacted by JYOTI. A human brain cDNA library was used. Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Baek JH., Mahon PC., Oh J., Kelly B., Krishnamachary B., Pearson M., Chan DA., Giaccia AJ., Semenza GL. Apoptosis - Interactions involving proteins with a function related to apoptosis gsemenza@jhmi.edu Accepted 2006-Not a month!-18 by DAVET. 23-JAN-2007: Contacted by JYOTI. Mol. Cell (1097-2765) 2005 Only protein-protein interactions imex curation egln3_human Prolyl hydroxylase EGLN3 Hypoxia-inducible factor prolyl hydroxylase 3 HPH-1 Prolyl hydroxylase domain-containing protein 3 EGLN3 Egl nine homolog 3 protein protein human Homo sapiens Human MPLGHIMRLDLEKIALEYIVPCLHEVGFCYLDNFLGEVVGDCVLERVKQLHCTGALRDGQLAGPRAGVSKRHLRGDQITWIGGNEEGCEAISFLLSLIDRLVLYCGSRLGKYYVKERSKAMVACYPGNGTGYVRHVDNPNGDGRCITCIYYLNKNWDAKLHGGILRIFPEGKSFIADVEPIFDRLLFFWSDRRNPHEVQPSYATRYAMTVWYFDAEERAEAKKKFRNLTRKTESALTED 9DA3A0F80168557B hif1a_human Hypoxia-inducible factor 1-alpha ARNT-interacting protein Member of PAS protein 1 Basic-helix-loop-helix-PAS protein MOP1 HIF1A MOP1 BHLHE78 PASD8 PAS domain-containing protein 8 Class E basic helix-loop-helix protein 78 protein protein human Homo sapiens Human MEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNCFYLKALDGFVMVLTDDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGLVKKGKEQNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKLKKEPDALTLLAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPNEKLQNINLAMSPLPTAETPKPLRSSADPALNQEVALKLEPNPESLELSFTMPQIQDQTPSPSDGSTRQSSPEPNSPSEYCFYVDSDMVNEFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESSSASPESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDIKILIASPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNVLSVALSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIGTLLQQPDDHAATTSLSWKRVKGCKSSEQNGMEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN DIP protein Q16665 original sequence version: 94 DIP protein Q16665 original sequence version: 98 DIP protein Q16665-1 original sequence version: 102 DIP protein Q16665 original sequence version: 102 DIP protein Q16665 original sequence version: 127 DIP protein Q16665 original sequence version: 120 DIP protein Q16665 original sequence version: 169 DIP protein Q16665 original sequence version: 174 DIP protein Q16665-1 original sequence version: 174 DIP protein Q16665-1 original sequence version: 191 DIP protein Q16665 original sequence version: 191 DIP protein Q16665 original sequence version: 179 DIP protein Q16665 original sequence version: 150 DIP protein Q16665 original sequence version: 156 DIP protein Q16665 original sequence version: 177 ABD4F7DAA135BE2D q13438-2 Protein OS-9 OS-9-2 OS9 Amplified in osteosarcoma 9 protein protein human Homo sapiens Human MAAETLLSSLLGLLLLGLLLPASLTGGVGSLNLEELSEMRYGIEILPLPVMGGQSQSSDVVIVSSKYKQRYECRLPAGAIHFQREREEETPAYQGPGIPELLSPMRDAPCLLKTKDWWTYEFCYGRHIQQYHMEDSEIKGEVLYLGYYQSAFDWDDETAKASKQHRLKRYHSQTYGNGSKCDLNGRPREAEVRFLCDEGAGISGDYIDRVDEPLSCSYVLTIRTPRLCPHPLLRPPPSAAPQAILCHPSLQPEEYMAYVQRQADSKQYGDKIIEELQDLGPQVWSETKSGVAPQKMAGASPTKDDSKDSDFWKMLNEPEDQAPGGEEVPAEEQDPSPEAADSASGAPNDFQNNVQVKVIRSPADLIRFIEELKGGTKKGKPNIGQEQPVDDAAEVPQREPEKERGDPERQREMEEEEDEDEDEDEDEDERQLLGEFEKELEGILLPSDRDRLRSEVKAGMERELENIIQETEKELDPDGLKKESERDRAMLALTSTLNKLIKRLEEKQSPELVKKHKKKRVVPKKPPPSPQPTGKIEIKIVRPWAEGTEEGARWLTDEDTRNLKEIFFNILVPGAEEAQKERQRQKELESNYRRVWGSPGGEGTGDLDEFDF Major isoform. 1FD06F7B23101015 q13438-1 Protein OS-9 OS-9-1 OS9 Amplified in osteosarcoma 9 protein protein human Homo sapiens Human MAAETLLSSLLGLLLLGLLLPASLTGGVGSLNLEELSEMRYGIEILPLPVMGGQSQSSDVVIVSSKYKQRYECRLPAGAIHFQREREEETPAYQGPGIPELLSPMRDAPCLLKTKDWWTYEFCYGRHIQQYHMEDSEIKGEVLYLGYYQSAFDWDDETAKASKQHRLKRYHSQTYGNGSKCDLNGRPREAEVRFLCDEGAGISGDYIDRVDEPLSCSYVLTIRTPRLCPHPLLRPPPSAAPQAILCHPSLQPEEYMAYVQRQADSKQYGDKIIEELQDLGPQVWSETKSGVAPQKMAGASPTKDDSKDSDFWKMLNEPEDQAPGGEEVPAEEQDPSPEAADSASGAPNDFQNNVQVKVIRSPADLIRFIEELKGGTKKGKPNIGQEQPVDDAAEVPQREPEKERGDPERQREMEEEEDEDEDEDEDEDERQLLGEFEKELEGILLPSDRDRLRSEVKAGMERELENIIQETEKELDPDGLKKESERDRAMLALTSTLNKLIKRLEEKQSPELVKKHKKKRVVPKKPPPSPQPTEEDPEHRVRVRVTKLRLGGPNQDLTVLEMKRENPQLKQIEGLVKELLEREGLTAAGKIEIKIVRPWAEGTEEGARWLTDEDTRNLKEIFFNILVPGAEEAQKERQRQKELESNYRRVWGSPGGEGTGDLDEFDF Major isoform. 65BA3F66CEC58756 egln1_human Egl nine homolog 1 EGLN1 C1orf12 PNAS-118 PNAS-137 Hypoxia-inducible factor prolyl hydroxylase 2 Prolyl hydroxylase domain-containing protein 2 SM-20 protein protein human Homo sapiens Human MANDSGGPGGPSPSERDRQYCELCGKMENLLRCSRCRSSFYCCKEHQRQDWKKHKLVCQGSEGALGHGVGPHQHSGPAPPAAVPPPRAGAREPRKAAARRDNASGDAAKGKVKAKPPADPAAAASPCRAAAGGQGSAVAAEAEPGKEEPPARSSLFQEKANLYPPSNTPGDALSPGGGLRPNGQTKPLPALKLALEYIVPCMNKHGICVVDDFLGKETGQQIGDEVRALHDTGKFTDGQLVSQKSDSSKDIRGDKITWIEGKEPGCETIGLLMSSMDDLIRHCNGKLGSYKINGRTKAMVACYPGNGTGYVRHVDNPNGDGRCVTCIYYLNKDWDAKVSGGILRIFPEGKAQFADIEPKFDRLLFFWSDRRNPHEVQPAYATRYAITVWYFDADERARAKVKYLTGEKGVRVELNKPSDSVGKDVF 81A97FF772CAA14C os9_human Protein OS-9 OS9 Amplified in osteosarcoma 9 protein protein human Homo sapiens Human MAAETLLSSLLGLLLLGLLLPASLTGGVGSLNLEELSEMRYGIEILPLPVMGGQSQSSDVVIVSSKYKQRYECRLPAGAIHFQREREEETPAYQGPGIPELLSPMRDAPCLLKTKDWWTYEFCYGRHIQQYHMEDSEIKGEVLYLGYYQSAFDWDDETAKASKQHRLKRYHSQTYGNGSKCDLNGRPREAEVRFLCDEGAGISGDYIDRVDEPLSCSYVLTIRTPRLCPHPLLRPPPSAAPQAILCHPSLQPEEYMAYVQRQADSKQYGDKIIEELQDLGPQVWSETKSGVAPQKMAGASPTKDDSKDSDFWKMLNEPEDQAPGGEEVPAEEQDPSPEAADSASGAPNDFQNNVQVKVIRSPADLIRFIEELKGGTKKGKPNIGQEQPVDDAAEVPQREPEKERGDPERQREMEEEEDEDEDEDEDEDERQLLGEFEKELEGILLPSDRDRLRSEVKAGMERELENIIQETEKELDPDGLKKESERDRAMLALTSTLNKLIKRLEEKQSPELVKKHKKKRVVPKKPPPSPQPTEEDPEHRVRVRVTKLRLGGPNQDLTVLEMKRENPQLKQIEGLVKELLEREGLTAAGKIEIKIVRPWAEGTEEGARWLTDEDTRNLKEIFFNILVPGAEEAQKERQRQKELESNYRRVWGSPGGEGTGDLDEFDF DIP protein Q13438 original sequence version: 63 DIP protein Q13438 original sequence version: 79 DIP protein Q13438-1 original sequence version: 136 65BA3F66CEC58756 vhl_human von Hippel-Lindau disease tumor suppressor VHL pVHL Protein G7 protein protein human Homo sapiens Human MPRRAENWDEAEVGAEEAGVEEYGPEEDGGEESGAEESGPEESGPEELGAEEEMEAGRPRPVLRSVNSREPSQVIFCNRSPRVVLPVWLNFDGEPQPYPTLPPGTGRRIHSYRGHLWLFRDAGTHDGLLVNQTELFVPSLNVDGQPIFANITLPVYTLKERCLQVVRSLVKPENYRRLDIVRSLYEDLEDHPNVQKDLERLTQERIAHQRMGD DIP protein P40337 original sequence version: 176 DIP protein P40337 original sequence version: 103 DIP protein P40337-1 original sequence version: 177 DIP protein P40337 original sequence version: 160 DIP protein P40337 original sequence version: 175 BA5D6765FBC16EA7 hif1a-os9 1 10 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 1 A The prey vectors started at nucleotides 484 and 1106 of the full length OS-9 mRNA sequence (GenBank accession number AB002806) and encoded fusion proteins containing aa 49-667 and 357-667, respectively, of OS-9 hif1a-os9-1 4 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 1 B HIF-1alpha residues 531-826, 653-826, or 692-826 efficiently bound to OS-9, whereas residues 1-329, 429-608, 531-610, and 786-826 did not bind OS-9 hif1a-os9-2 4 7 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1174659]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 667-667, sequence length 612 [EBI-1174659]589-667 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 1 C GST-HIF-1alpha(531-826) bound to in vitro-translated OS-9 isoform 1 (aa 1-667) ing OS-9 or PHD2. PHD2 was chosen for analysis beand isoform 2 (aa 1-533/589-667) hif1a-os9-3 4 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction 1 C Testing of carboxy-terminal truncation mutants of isoform 1 in the GST pull-down assay demonstrated binding of OS-9 aa 1-544 to HIF-1alpha, whereas OS-9 aa 1-235 and 1-158 did not bind. os9-egln1 2 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 A endogenous PHD2 coimmunoprecipitated with V5-OS-9 and FLAG-HIF-1alpha When the cells were not transfected with FLAG-HIF-1-alpha, V5-OS-9 immunoprecipiated PHD2. Q13438 transfection os9-egln1-1 2 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey association association 2 A endogenous PHD2 coimmunoprecipitated with V5-OS-9 and FLAG-HIF-1alpha These interactions took place in hypoxic (1% O2) or nonhypoxic (20% O2) conditions. Q16665 overexpressed Q13438; Q16665 transfection os9-hif1a 2 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 B Q13438 transfection cells were transfected with V5-OS-9 ex- pression vector and treated with desferrioxamine (DFO) to induce endogenous HIF-1alpha expression. Coimmuno- precipitation was demonstrated only in cells expressing both V5-OS-9 and endogenous HIF-1alpha Desferrioxamine (DFO) was used to induce endogenous HIF-1alpha expression. os9-egln1-2 2 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey association association 3 D Q13438 transfection V5-OS-9 coimmunoprecipitated HIF-1alpha-TM as efficiently as native HIF-1alpha These interactions took place in the presence of desferrioxamine which induces the expression of endogenous HIF-1-alpha. os9-egln1-3 2 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey Q16665:p.[Pro402Ala;Pro564Ala;Asn803Ala] mutation mutation alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain P A certain certain sequence position certain certain certain sequence position certain P A certain certain sequence position certain certain certain sequence position certain N A Wed May 02 16:52:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro402ala,pro564ala,asn803ala' Tue Jun 30 17:56:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Pro402Ala;Pro564Ala;Asn803Ala]' 9 unspecified role unspecified role prey prey association association 3 D Q13438; Q16665 transfection V5-OS-9 coimmunoprecipitated HIF-1alpha-TM as efficiently as native HIF-1alpha These interactions took place in the presence of desferrioxamine which induces the expression of endogenous HIF-1 alpha. The triple mutant of HIF-1 alpha was coimmunoprecipitated by V5-OS-9 as efficiently as the native HIF-1 alpha. os9-egln3 2 10 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey physical association physical association 5 A Q13438 transfection PHD3 also interacted with V5-OS-9 in 293 cells os9-egln3-1 4 10 unspecified role unspecified role prey prey region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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. 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role prey prey 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. association association 5 B when 293 cell lysates were incubated with GST-HIF-1alpha(531-826) in vitro, the binding of PHD3 was dramatically increased by coexpression of V5-OS-9 The binding of PHD3 to HIF-1 was increased by coexpression of V5-OS-9. hif1a-os9-4 4 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 10 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) association association 6 A Although OS-9 bound to either hydroxylated or nonhydroxylated GST-HIF-1alpha, increasing amounts of OS-9 had no effect on the binding of VHL to hydroxylated GST-HIF-1alpha Increasing amounts of OS-9 did not have an effect on the binding of VHL to hydroxylated GST-HIF-1-alpha. The GST-HIF-1-alpha was incubated with rabbit reticulolysate to promote hydroxylation. The VHL did not interact with the non-hydroxylated HIF-1-alpha. hif1a-os9-5 4 6 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 10 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 6 A OS-9 bound to either hydroxylated or nonhydroxylated GST-HIF-1alpha hif1a-vhl 4 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) physical association physical association 6 A increasing amounts of OS-9 had no effect on the binding of VHL to hydroxylated to GST-HIF-1alpha This interaction takes place in the absence of V5-OS-9 and in the presence of hydroxylated HIF-1-alpha. hif1a-vhl-1 3 11 unspecified role unspecified role prey prey 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. 11 unspecified role unspecified role prey prey region-x 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role bait bait region-x gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal-x sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) association association 6 A increasing amounts of OS-9 had no effect on the binding of VHL to hydroxylated to GST-HIF-1alpha This interaction takes place in the absence of V5-OS-9 and in the presence of hydroxylated HIF-1-alpha. The binding of VHL was greatly increased following incubation of GST-HIF-1alpha(531-826) with lysates from PHD2-transfected cells. VHL binding was further increased when lysates from cells cotransfected with both PHD2 and OS-9 were used. Addition of dimethyloxalylglycine (DMOG), a competitive antagonist of 2-oxoglutarate, blocked binding of VHL to GST-HIF-1alpha(531-826), even when lysates containing both PHD2 and OS-9 were used 6 B