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/ zhou-2004b-5 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. mouse-l cells Mus musculus embryo fibroblast, Moloney Sarcoma Virus transformed l cells Mouse embryo fibroblast, Moloney Sarcoma Virus transformed anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 zhou-2004b-6 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. 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 predetermined predetermined participant predetermined Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 zhou-2004b-1 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. mouse-l cells Mus musculus embryo fibroblast, Moloney Sarcoma Virus transformed l cells Mouse embryo fibroblast, Moloney Sarcoma Virus transformed confocal microscopy confocal microscopy immunostaining immunostaining Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 zhou-2004b-2 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. rat-mca_rh7777 Rattus norvegicus McA-RH7777 cells (hepatoma) mca_rh7777 buffalo rat hepatoma cells Species: rat, buffalo female; Tissue: liver; Tumor: hepatoma; Derived from: Morris hepatoma 7777 confocal microscopy confocal microscopy immunostaining immunostaining Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 zhou-2004b-3 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. mouse-l cells Mus musculus embryo fibroblast, Moloney Sarcoma Virus transformed l cells Mouse embryo fibroblast, Moloney Sarcoma Virus transformed fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. The species of the SCP-2 protein was obtained by looking at a reference from one of the papers referenced (Seedorf et al. (1994) J Bio Chem 269 2613). The species of the caveolin-1 protein was inferred by looking at a publication by the donor of the construct. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 zhou-2004b-4 Sterol carrier protein-2 directly interacts with caveolin-1 in vitro and in vivo. mouse-l cells Mus musculus embryo fibroblast, Moloney Sarcoma Virus transformed l cells Mouse embryo fibroblast, Moloney Sarcoma Virus transformed electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy immunostaining immunostaining Zhou M., Parr RD., Petrescu AD., Payne HR., Atshaves BP., Kier AB., Ball JM., Schroeder F. Cancer - Interactions investigated in the context of cancer Mock-transfected cells were double immunolabeled with anti-sera to SCP-2 and caveolin- 1 followed by reaction with secondary antibodies conjugated to Rhodamine Red X and FITC, respectively. The cells were examined by laser scanning confocal microscopy. fschroeder@cvm.tamu.edu Biochemistry (0006-2960) 2004 cav1_mouse Caveolin-1 Cav1 Cav protein protein mouse Mus musculus Mouse MSGGKYVDSEGHLYTVPIREQGNIYKPNNKAMADEVTEKQVYDAHTKEIDLVNRDPKHLNDDVVKIDFEDVIAEPEGTHSFDGIWKASFTTFTVTKYWFYRLLSTIFGIPMALIWGIYFAILSFLHIWAVVPCIKSFLIEIQCISRVYSIYVHTFCDPLFEAIGKIFSNIRISTQKEI scp2_human Sterol carrier protein 2 Propanoyl-CoA C-acyltransferase Sterol carrier protein X SCP-chi SCPX SCP2 Acetyl-CoA C-myristoyltransferase Non-specific lipid-transfer protein SCP-2/3-oxoacyl-CoA thiolase SCP-2/thiolase Straight-chain acyl-CoA oxidase protein protein human Homo sapiens Human MSSSPWEPATLRRVFVVGVGMTKFVKPGAENSRDYPDLAEEAGKKALADAQIPYSAVDQACVGYVFGDSTCGQRAIYHSLGMTGIPIINVNNNCATGSTALFMARQLIQGGVAECVLALGFEKMSKGSLGIKFSDRTIPTDKHVDLLINKYGLSAHPVAPQMFGYAGKEHMEKYGTKIEHFAKIGWKNHKHSVNNPYSQFQDEYSLDEVMASKEVFDFLTILQCCPTSDGAAAAILASEAFVQKYGLQSKAVEILAQEMMTDLPSSFEEKSIIKMVGFDMSKEAARKCYEKSGLTPNDIDVIELHDCFSTNELLTYEALGLCPEGQGATLVDRGDNTYGGKWVINPSGGLISKGHPLGATGLAQCAELCWQLRGEAGKRQVPGAKVALQHNLGIGGAVVVTLYKMGFPEAASSFRTHQIEAVPTSSASDGFKANLVFKEIEKKLEEEGEQFVKKIGGIFAFKVKDGPGGKEATWVVDVKNGKGSVLPNSDKKADCTITMADSDFLALMTGKMNPQSAFFQGKLKITGNMGLAMKLQNLQLQPGNAKL cav1_human Caveolin-1 CAV1 CAV protein protein human Homo sapiens Human MSGGKYVDSEGHLYTVPIREQGNIYKPNNKAMADELSEKQVYDAHTKEIDLVNRDPKHLNDDVVKIDFEDVIAEPEGTHSFDGIWKASFTTFTVTKYWFYRLLSALFGIPMALIWGIYFAILSFLHIWAVVPCIKSFLIEIQCISRVYSIYVHTVCDPLFEAVGKIFSNVRINLQKEI scp2_mouse Sterol carrier protein 2 Scp2 Scp-2 Propanoyl-CoA C-acyltransferase Sterol carrier protein X SCP-chi Acetyl-CoA C-myristoyltransferase Non-specific lipid-transfer protein SCP-2/3-oxoacyl-CoA thiolase SCP-2/thiolase Straight-chain acyl-CoA oxidase protein protein mouse Mus musculus Mouse MPSVALKSPRLRRVFVVGVGMTKFMKPGGENSRDYPDMAKEAGQKALEDAQIPYSAVEQACVGYVYGDSTSGQRAIYHSLGLTGIPIINVNNNCSTGSTALFMAHQLIQGGLANCVLALGFEKMERGSIGTKFSDRTTPTDKHIEVLIDKYGLSAHPITPQMFGYAGKEHMEKYGTKVEHFAKIGWKNHKHSVNNTYSQFQDEYSLEEVMKSKPVFDFLTILQCCPTSDGAAAAILSSEEFVQQYGLQSKAVEIVAQEMMTDLPSTFEEKSIIKVVGYDMSKEAARRCYEKSGLTPNDVDVIELHDCFSVNELITYEALGLCPEGQGGTLVDRGDNTYGGKWVINPSGGLISKGHPLGATGLAQCAELCWQLRGEAGKRQVPGAKVALQHNLGLGGAVVVTLYRMGFPEAASSFRTHQVSAAPTSSAGDGFKANLVFKEIEKKLEEEGEQFVKKIGGIFAFKVKDGPGGKEATWVVDVKNGKGSVLPNSDKKADCTITMADSDLLALMTGKMNPQSAFFQGKLKIAGNMGLAMKLQNLQLQPGKAKL scp2_rat Sterol carrier protein 2 Propanoyl-CoA C-acyltransferase Sterol carrier protein X SCP-chi Scp2 Scp-2 Acetyl-CoA C-myristoyltransferase Non-specific lipid-transfer protein SCP-2/3-oxoacyl-CoA thiolase SCP-2/thiolase Straight-chain acyl-CoA oxidase protein protein rat Rattus norvegicus (Rat) Rat MPSVALNSPRLPRVFVVGVGMTKFMKPGGENSRDYPDLAKEAGQKALADRQIPYSAVEQACVGYVYGESTCGQRAIYHSLGLTGIPIINVNNNCSTGSTALFMAQQLVQGGLANCVLALGFEKMEKGSLGTKYSDRSNPLEKHIDVLINKYGMSACPFAPQLFGSAGKEHMETYGTKVEHFAKIGWKNHKHSVNNPYSQFQDEYSLDEIMKSRPVFDFLTVLQCCPTSDGAAAAIVSSEEFVQKHGLQSKAVEIVAQEMVTDMPSTFEEKSVIKMVGYDMSKEAARKCYEKSGLGPSDVDVIELHDCFSTNELLTYEALGLCPEGQGGALVDRGDNTYGGKWVINPSGGLISKGHPLGATGLAQCAELCWQLRGEAGKRQVPGAKVALQHNLGLGGAAVVTLYRMGFPEAASSFRTHQISAAPTSSAGDGFKANLIFKEIEKKLEEEGEEFVKKIGGIFAFKVKDGPGGKEATWVVDVKNGKGSVLPDSDKKADCTITMADSDLLALMTGKMNPQSAFFQGKLKIAGNMGLAMKLQSLQLQPDKAKL cav1_rat Caveolin-1 Cav1 Cav protein protein rat Rattus norvegicus (Rat) Rat MSGGKYVDSEGHLYTVPIREQGNIYKPNNKAMADEVNEKQVYDAHTKEIDLVNRDPKHLNDDVVKIDFEDVIAEPEGTHSFDGIWKASFTTFTVTKYWFYRLLSTIFGIPMALIWGIYFAILSFLHIWAVVPCIKSFLIEIQCISRVYSIYVHTFCDPLFEAIGKIFSNIRISTQEEI scp2-cav1-2 Anti-SCP-2 antibody coprecipitated caveolin-1 567416 N/A 567422 unspecified role unspecified role prey prey N/A 567423 unspecified role unspecified role bait bait physical association physical association false false false The L-cells overexpressed human SCP-2. 8 cav1-scp2-2 Anti-caveolin-1 antibody coprecipitated SCP-2 567416 N/A 567423 unspecified role unspecified role prey prey N/A 567422 unspecified role unspecified role bait bait physical association physical association false false false The L-cells overexpressed human SCP-2. 8 scp2-cav1-3 567417 N/A 567424 unspecified role unspecified role prey prey N/A 567423 unspecified role unspecified role bait bait physical association physical association false false false 12, Table 2 cav1-scp2-3 567417 N/A 567423 unspecified role unspecified role prey prey N/A 567424 unspecified role unspecified role bait bait physical association physical association false false false 12, Table 2 cav1-scp2 567418 N/A 567425 unspecified role unspecified role neutral component neutral component N/A 567422 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 1 H, Table 1 In L-cell plasma membranes, 86.7 plus or minus 9.9% of SCP-2-pixels were colocalized with 87.6 plus or minus 10.9% of caveolin-1-pixels. Plasma membranes of cells containing little SCP-2 but much caveolin-1 (i.e., L-cells) exhibited 3.5-fold more colocalized SCP-2/caveolin-1 than plasma membranes of cells containing high levels of SCP-2 but relatively little caveolin-1 (i.e., hepatoma cells). However, in both cases at least part of SCP-2 was structurally colocalized with caveolin- 1, primarily at the plasma membrane in L-cells, and more so in punctate (vesicles) structures containing caveolin-1 in hepatoma cells. cav1-scp2-1 567419 N/A 567426 unspecified role unspecified role neutral component neutral component N/A 567427 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 1 D, Table 1 Quantitative pixel fluorogram analysis within select plasma membrane regions showed that an average of 24.8 plus or minus 13.4% of SCP-2 -pixels were colocalized with 75.6 plus or minus 15.3% of caveolin-1-pixels within the hepatoma cell plasma membrane. Plasma membranes of cells containing little SCP-2 but much caveolin-1 (i.e., L-cells) exhibited 3.5-fold more colocalized SCP-2/caveolin-1 than plasma membranes of cells containing high levels of SCP-2 but relatively little caveolin-1 (i.e., hepatoma cells). However, in both cases at least part of SCP-2 was structurally colocalized with caveolin- 1, primarily at the plasma membrane in L-cells, and more so in punctate (vesicles) structures containing caveolin-1 in hepatoma cells. scp2-cav1 FRET indicates SCP-2 colocalizes with Caveolin-1 in L-cells 567420 N/A 567423 unspecified role unspecified role fluorescence donor fluorescence donor region Cy3 donor tag cy3 label cy3 label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 567424 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region Cy5 acceptor tag cy5 label cy5 label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false proximity proximity close-contact colocalization neighbourhood interaction false false false 2 scp2-cav1-1 Immunogold electron microscopy indicates SCP-2 and caveolin-1 colocalize 567421 N/A 567423 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 567422 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 colocalization colocalization false false false Human SCP-2 was overexpressed in L-cells. 3 A, B The data indicate SCP-2 and caveolin-1 colocalize within the plasma membrane in regions well within the size of caveolae. The distance between the proteins as determined by immunogold electron microscopy is greater than for FRET as the colloidal gold particles are larger than Cy3/Cy5, there is repulsion between the gold particles and fewer measurements were taken.