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/ szczesna_s-2011-1 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation szczesna_s-2011-3 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation szczesna_s-2011-5 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation szczesna_s-2011-2 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation szczesna_s-2011-7 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation szczesna_s-2011-4 Progesterone receptor membrane component 1 inhibits the activity of drug-metabolizing cytochromes P450 and binds to cytochrome P450 reductase. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Szczesna-Skorupa E., Kemper B. Cancer - Interactions investigated in the context of cancer There has been no clarification from the authors regarding the construct "C1(1–29)/GFP":- notably if it is derived from CYP2C2 (as stated in this publication in the results section) or from CYP2C1, as given in the quoted reference:- Szczesna-Skorupa et al., 1998. (There is a 3 amino acid difference between the two sequences.) In figure 8B participants were subsequently identified by molecular weight. C1(1–29)/GFP was found to contain the first 29 amino acids from Rabbit CYP2C1 - Ahn et al,. JBC 1993. OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide. byronkem@life.illinois.edu Mol. Pharmacol. (0026-895X) 2011 Only protein-protein interactions imex curation pgrc1_human Membrane-associated progesterone receptor component 1 Dap1 IZA PGRMC1 HPR6.6 PGRMC protein protein human Homo sapiens Human MAAEDVVATGADPSDLESGGLLHEIFTSPLNLLLLGLCIFLLYKIVRGDQPAASGDSDDDEPPPLPRLKRRDFTPAELRRFDGVQDPRILMAINGKVFDVTKGRKFYGPEGPYGVFAGRDASRGLATFCLDKEALKDEYDDLSDLTAAQQETLSDWESQFTFKYHHVGKLLKEGEEPTVYSDEEEPKDESARKND cp2c2_rabit Cytochrome P450 2C2 CYP2C2 CYPIIC2 Cytochrome P450 PBc2 Cytochrome P450 PHP2 Laurate omega-1 hydroxylase protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MDLVVVLGLCLSCLLLPSLWKQSHGGGKLPPGPTPFPILGNVLQLDFKDLSKSLTNLSKVYGPVFTVYLGMKPTVVVHGYEAVKEALVDLGHELSGRSRFLVTAKLNKGFGVIFSNGKRWTETRRFSLMTLRNFGMGKRSIEERVQEEAHCLVEELRKTNASPCDPTFILGAAPCNVICSVIFQNRFDYTDQDFLSLMGKFNENFKILNSPWVQFCNCFPILFDYFPGSHRKAVKNIFYVKNYITEQIKEHQKSLDINNPRDFIDCFLIKMEQEKCNQQSEFTIENLLTTVSDVFMAGTETTSTTLRYGLLLLMKHPEVIAKVQEEIERVIGRHRSPCMQDRSRMPYTDATVHEIQRYINLIPNNVPHTTICNLKFRNYLIPKGTDVLTSLSSVLHDDKEFPNPDRFDPGHFLDASGNFRKSDYFMPFSTGKRVCVGEALARMELFLFLTAILQNFTPKPLVNPNNVDENPFSSGIVRVPPLYRVSFIPV cp2c8_human Cytochrome P450 2C8 CYP2C8 CYPIIC8 Cytochrome P450 IIC2 Cytochrome P450 MP-12 Cytochrome P450 MP-20 Cytochrome P450 form 1 S-mephenytoin 4-hydroxylase protein protein human Homo sapiens Human MEPFVVLVLCLSFMLLFSLWRQSCRRRKLPPGPTPLPIIGNMLQIDVKDICKSFTNFSKVYGPVFTVYFGMNPIVVFHGYEAVKEALIDNGEEFSGRGNSPISQRITKGLGIISSNGKRWKEIRRFSLTTLRNFGMGKRSIEDRVQEEAHCLVEELRKTKASPCDPTFILGCAPCNVICSVVFQKRFDYKDQNFLTLMKRFNENFRILNSPWIQVCNNFPLLIDCFPGTHNKVLKNVALTRSYIREKVKEHQASLDVNNPRDFIDCFLIKMEQEKDNQKSEFNIENLVGTVADLFVAGTETTSTTLRYGLLLLLKHPEVTAKVQEEIDHVIGRHRSPCMQDRSHMPYTDAVVHEIQRYSDLVPTGVPHAVTTDTKFRNYLIPKGTTIMALLTSVLHDDKEFPNPNIFDPGHFLDKNGNFKKSDYFMPFSAGKRICAGEGLARMELFLFLTTILQNFNLKSVDDLKNLNTTAVTKGIVSLPPSYQICFIPV cp2c1_rabit Cytochrome P450 2C1 CYP2C1 CYPIIC1 Cytochrome P-450 IIC1 Cytochrome P450 PBc1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MDPVVVLGLCLSCLLLLSLWKQSYGGGKLPPGPTPFPILGNILQIGIQDISKSFTKLSEVYGPVFTVYLGMKPTVVIHGYDAVKEALVDLGEEFSGRIVFPLTAKINKGYGIVFSNGKRWKETRRFSLMTLRDFGMGKRSIEDRVQEEARCLVEELRKTNGSPCNPTFILGAAPCNVICSVIFQNRFDYTDQDFLSLMGKLNENFKILNSPWVQMCNNFPILIDYLPGSHNKILRNNIYIRNYVLEKIKEHQETLDINNPRDFIDCFLIKMEQEKDNQQSEFTIENLMTTLSDVFGAGTETTSTTLRYGLLLLMKHPEVIAKVQEEIERVIGRHRSPCMQDRSRMPYTDATVHEIQRYINLIPNNVPRATTCNVKFRSYLIPKGTAVITSLTSMLYNDKEFPNPDRFDPGHFLDASGKFRKSDYFMPFSTGKRVCVGEVLARMELFLFLTAILQNFTPKPLVDPKDIDTTPLVSGLGRVPPLYQLSFIPA ncpr_human NADPH--cytochrome P450 reductase POR CYPOR protein protein human Homo sapiens Human MGDSHVDTSSTVSEAVAEEVSLFSMTDMILFSLIVGLLTYWFLFRKKKEEVPEFTKIQTLTSSVRESSFVEKMKKTGRNIIVFYGSQTGTAEEFANRLSKDAHRYGMRGMSADPEEYDLADLSSLPEIDNALVVFCMATYGEGDPTDNAQDFYDWLQETDVDLSGVKFAVFGLGNKTYEHFNAMGKYVDKRLEQLGAQRIFELGLGDDDGNLEEDFITWREQFWPAVCEHFGVEATGEESSIRQYELVVHTDIDAAKVYMGEMGRLKSYENQKPPFDAKNPFLAAVTTNRKLNQGTERHLMHLELDISDSKIRYESGDHVAVYPANDSALVNQLGKILGADLDVVMSLNNLDEESNKKHPFPCPTSYRTALTYYLDITNPPRTNVLYELAQYASEPSEQELLRKMASSSGEGKELYLSWVVEARRHILAILQDCPSLRPPIDHLCELLPRLQARYYSIASSSKVHPNSVHICAVVVEYETKAGRINKGVATNWLRAKEPAGENGGRALVPMFVRKSQFRLPFKATTPVIMVGPGTGVAPFIGFIQERAWLRQQGKEVGETLLYYGCRRSDEDYLYREELAQFHRDGALTQLNVAFSREQSHKVYVQHLLKQDREHLWKLIEGGAHIYVCGDARNMARDVQNTFYDIVAELGAMEHAQAVDYIKKLMTKGRYSLDVWS insi1_human Insulin-induced gene 1 protein INSIG1 protein protein human Homo sapiens Human MPRLHDHFWSCSCAHSARRRGPPRASAAGLAAKVGEMINVSVSGPSLLAAHGAPDADPAPRGRSAAMSGPEPGSPYPNTWHHRLLQRSLVLFSVGVVLALVLNLLQIQRNVTLFPEEVIATIFSSAWWVPPCCGTAAAVVGLLYPCIDSHLGEPHKFKREWASVMRCIAVFVGINHASAKLDFANNVQLSLTLAALSLGLWWTFDRSRSGLGLGITIAFLATLITQFLVYNGVYQYTSPDFLYIRSWLPCIFFSGGVTVGNIGRQLAMGVPEKPHSD cp3a4_human Cytochrome P450 3A4 Albendazole monooxygenase (sulfoxide-forming) CYP3A4 CYP3A3 Albendazole sulfoxidase CYPIIIA3 CYPIIIA4 Cytochrome P450 3A3 Cytochrome P450 HLp Cytochrome P450 NF-25 Cytochrome P450-PCN1 Nifedipine oxidase Quinine 3-monooxygenase 1,8-cineole 2-exo-monooxygenase Cholesterol 25-hydroxylase 1,4-cineole 2-exo-monooxygenase protein protein human Homo sapiens Human MALIPDLAMETWLLLAVSLVLLYLYGTHSHGLFKKLGIPGPTPLPFLGNILSYHKGFCMFDMECHKKYGKVWGFYDGQQPVLAITDPDMIKTVLVKECYSVFTNRRPFGPVGFMKSAISIAEDEEWKRLRSLLSPTFTSGKLKEMVPIIAQYGDVLVRNLRREAETGKPVTLKDVFGAYSMDVITSTSFGVNIDSLNNPQDPFVENTKKLLRFDFLDPFFLSITVFPFLIPILEVLNICVFPREVTNFLRKSVKRMKESRLEDTQKHRVDFLQLMIDSQNSKETESHKALSDLELVAQSIIFIFAGYETTSSVLSFIMYELATHPDVQQKLQEEIDAVLPNKAPPTYDTVLQMEYLDMVVNETLRLFPIAMRLERVCKKDVEINGMFIPKGVVVMIPSYALHRDPKYWTEPEKFLPERFSKKNKDNIDPYIYTPFGSGPRNCIGMRFALMNMKLALIRVLQNFSFKPCKETQIPLKLSLGGLLQPEKPVVLKVESRDGTVSGA bap31_human B-cell receptor-associated protein 31 BCAP31 BAP31 p28 6C6-AG tumor-associated antigen DXS1357E Protein CDM protein protein human Homo sapiens Human MSLQWTAVATFLYAEVFVVLLLCIPFISPKRWQKIFKSRLVELLVSYGNTFFVVLIVILVLLVIDAVREIRKYDDVTEKVNLQNNPGAMEHFHMKLFRAQRNLYIAGFSLLLSFLLRRLVTLISQQATLLASNEAFKKQAESASEAAKKYMEENDQLKKGAAVDGGKLDVGNAEVKLEEENRSLKADLQKLKDELASTKQKLEKAENQVLAMRKQSEGLTKEYDRLLEEHAKLQAAVDGPMDKKEE pgrmc1-cyp2c2 1308088 n/a 1308094 unspecified role unspecified role bait bait 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 false n/a 1308095 unspecified role unspecified role prey prey c_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 1A cyp2c8-cyp2c2 1308088 n/a 1308096 unspecified role unspecified role bait bait c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308094 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 false physical association physical association false false false Fig 1A pgrmc1-cyp2c2-1 1308088 n/a 1308097 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n_terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1308094 unspecified role unspecified role bait bait 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 false physical association physical association false false false Fig 1B the chimera OEC/GFP binds much more efficiently to PGRMC1 than C1(1–29)/GFP, consistent with an interaction mediated mainly by the cytosolic domain of CYP2C2 (Fig. 1B, left). It is possible that the construct "C1(1–29)/GFP" may have been derived from CYP2C2 rather than CYP2C1- as entered here. pgrmc1-cyp2c2 1308088 n/a 1308094 unspecified role unspecified role bait bait 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 false n/a 1308095 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cytoplasmic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false egfr_transmembrane_domain tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 1B the chimera OEC/GFP binds much more efficiently to PGRMC1 than C1(1–29)/GFP, consistent with an interaction mediated mainly by the cytosolic domain of CYP2C2 (Fig. 1B, left). pgrmc1-cyp2c2-3 1308088 n/a 1308095 unspecified role unspecified role prey prey c_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308098 unspecified role unspecified role prey prey c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308094 unspecified role unspecified role bait bait 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 false association association false false false Fig 8B In cells coexpressing all three proteins, there was a marked decrease in CPR binding to PGRMC1, whereas binding of PGRMC1 to CYP2C2 was little changed. pgrmc1-cyp2c2-2 1308088 n/a 1308095 unspecified role unspecified role prey prey c_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308094 unspecified role unspecified role bait bait 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 false physical association physical association false false false Fig 8B pgrmc1-cyp2c2-4 1308088 n/a 1308094 unspecified role unspecified role bait bait 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 false n/a 1308098 unspecified role unspecified role prey prey c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 8B por-cyp2c2-2 1308088 n/a 1308095 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 false 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 n/a 1308099 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 10A binding of INSIG1 to CYP2C2 is independent of PGRMC1. cyp2c2-insig1 1308088 n/a 1308095 unspecified role unspecified role bait bait 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 false 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 n/a 1308099 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 10A Binding of INSIG1 to CYP2C2 is independent of PGRMC1. pgrmc1-cyp2c1 1308088 n/a 1308099 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1308097 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n_terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 10C -1 Coimmunoprecipitation of INSIG1 and chimera C1(1–29)/GFP is more efficient than that with the cytoplasmic domain chimera OEC/GFP. [OEC is a fusion protein assembled from the catalytic cytosolic domain (amino acids 21–490) of CYP2C2 fused to the transmembrane domain of epidermal growth factor receptor (EGFR- species undetermined), -which replaces the deleted CYP2C2 signal peptide.] cyp2c1-insig1 1308088 n/a 1308099 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1308097 unspecified role unspecified role bait bait n_terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 10C -2 Coimmunoprecipitation of INSIG1 and chimera C1(1–29)/GFP is more efficient than that with the cytoplasmic domain chimera OEC/GFP. cyp2c2-insig1-1 1308088 n/a 1308099 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1308095 unspecified role unspecified role bait bait egfr_transmembrane_domain tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cytoplasmic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 10C-3 Coimmunoprecipitation of INSIG1 and chimera C1(1–29)/GFP is more efficient than that with the cytoplasmic domain chimera OEC/GFP (Fig. 10C). cyp2c2-pgrmc1 1308089 n/a 1308094 unspecified role unspecified role neutral component neutral component 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 false n/a 1308095 unspecified role unspecified role neutral component neutral component c_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig 2A cyp3a4-pgrmc1-2 1308089 n/a 1308094 unspecified role unspecified role neutral component neutral component 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 false n/a 1308100 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig 2A cyp3a4-pgrmc1-3 1308089 n/a 1308094 unspecified role unspecified role neutral component neutral component 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 false n/a 1308098 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig 9A cyp2c2-bcap31 1308090 n/a 1308101 unspecified role unspecified role prey prey n/a 1308095 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 1B pgrmc1-por 1308090 n/a 1308098 unspecified role unspecified role prey prey c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308094 unspecified role unspecified role bait bait 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 false physical association physical association false false false Fig 8A cyp2c2-por 1308090 n/a 1308095 unspecified role unspecified role bait bait 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 false 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 n/a 1308098 unspecified role unspecified role prey prey c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 8A cyp3a4-pgrmc1 1308091 n/a 1308094 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 false n/a 1308100 unspecified role unspecified role bait bait region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 1A por-cyp2c2-3 1308091 n/a 1308098 unspecified role unspecified role bait bait c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308095 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 false 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 physical association physical association false false false Fig 10B por-cyp2c2 1308092 n/a 1308094 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 false n/a 1308098 unspecified role unspecified role bait bait c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1308095 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 false 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 association association false false false Fig 8C -1 only weak binding of PGRMC1 to CPR was observed (Fig. 8C, third panel). por-cyp2c2-1 1308092 n/a 1308095 unspecified role unspecified role prey prey 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 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 false n/a 1308098 unspecified role unspecified role bait bait c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 8C -2 In cells coexpressing all three proteins, there was a marked decrease in CPR binding to PGRMC1, whereas binding of PGRMC1 to CYP2C2 was little changed. cyp2c2-pgrmc1-1 1308093 n/a 1308094 unspecified role unspecified role neutral component neutral component 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 false n/a 1308095 unspecified role unspecified role neutral component neutral component c_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig 2A/B