UniProt UniProt http://www.uniprot.org ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. Biochimica et biophysica acta 2012 imex curation Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Only protein-protein interactions imex curation human-huh7 Homo sapiens hepatoma cell line established from a hepatocellular carcinoma huh7 Human hepatoma cell line established from a hepatocellular carcinoma bifc bimolecular fluorescence complementation bifc anti tag western anti tag western blot anti tag western Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Accepted 2020-JUN-15 AT 11:19 BST AT 11:19 BST by MGT Biochimica et biophysica acta 2012 Only protein-protein interactions imex curation imex curation ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. Biochimica et biophysica acta 2012 imex curation Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Only protein-protein interactions imex curation human-huh7 Homo sapiens hepatoma cell line established from a hepatocellular carcinoma huh7 Human hepatoma cell line established from a hepatocellular carcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Accepted 2020-JUN-15 AT 11:19 BST AT 11:19 BST by MGT Biochimica et biophysica acta 2012 Only protein-protein interactions imex curation imex curation ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. Biochimica et biophysica acta 2012 imex curation Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Only protein-protein interactions imex curation in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay antibody detection identification by antibody antibody detection Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Accepted 2020-JUN-15 AT 11:19 BST AT 11:19 BST by MGT Biochimica et biophysica acta 2012 Only protein-protein interactions imex curation imex curation ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. ORP10, a cholesterol binding protein associated with microtubules, regulates apolipoprotein B-100 secretion. Biochimica et biophysica acta 2012 imex curation Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. vesa.olkkonen@helsinki.fi Only protein-protein interactions imex curation in vitro In vitro affinity techniques affinity technology affinity techniques tag visualisation tag visualisation Nissilä E., Ohsaki Y., Weber-Boyvat M., Perttilä J., Ikonen E., Olkkonen VM. Cholesterol binding was assayed by measuring the ability of GST-ORP10 to extract [3H] cholesterol from large unilamellar PC-PE vesicles containing 1 mol% cholesterol. vesa.olkkonen@helsinki.fi Accepted 2020-JUN-15 AT 11:19 BST AT 11:19 BST by MGT Biochimica et biophysica acta 2012 Only protein-protein interactions imex curation imex curation phosphatidic acids phosphatidic acids small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) osbl9_human Oxysterol-binding protein-related protein 9 OSBPL9 ORP9 OSBP4 protein protein human Homo sapiens Human MASIMEGPLSKWTNVMKGWQYRWFVLDYNAGLLSYYTSKDKMMRGSRRGCVRLRGAVIGIDDEDDSTFTITVDQKTFHFQARDADEREKWIHALEETILRHTLQLQGLDSGFVPSVQDFDKKLTEADAYLQILIEQLKLFDDKLQNCKEDEQRKKIETLKETTNSMVESIKHCIVLLQIAKDQSNAEKHADGMISTINPVDAIYQPSPLEPVISTMPSQTVLPPEPVQLCKSEQRPSSLPVGPVLATLGHHQTPTPNSTGSGHSPPSSSLTSPSHVNLSPNTVPEFSYSSSEDEFYDADEFHQSGSSPKRLIDSSGSASVLTHSSSGNSLKRPDTTESLNSSLSNGTSDADLFDSHDDRDDDAEAGSVEEHKSVIMHLLSQVRLGMDLTKVVLPTFILERRSLLEMYADFFAHPDLFVSISDQKDPKDRMVQVVKWYLSAFHAGRKGSVAKKPYNPILGEIFQCHWTLPNDTEENTELVSEGPVPWVSKNSVTFVAEQVSHHPPISAFYAECFNKKIQFNAHIWTKSKFLGMSIGVHNIGQGCVSCLDYDEHYILTFPNGYGRSILTVPWVELGGECNINCSKTGYSANIIFHTKPFYGGKKHRITAEIFSPNDKKSFCSIEGEWNGVMYAKYATGENTVFVDTKKLPIIKKKVRKLEDQNEYESRSLWKDVTFNLKIRDIDAATEAKHRLEERQRAEARERKEKEIQWETRLFHEDGECWVYDEPLLKRLGAAKH A689D5D68E95EC6C osb11_human Oxysterol-binding protein-related protein 11 OSBPL11 ORP11 OSBP12 protein protein human Homo sapiens Human MQGGEPVSTMKVSESEGKLEGQATAVTPNKNSSCGGGISSSSSSRGGSAKGWQYSDHMENVYGYLMKYTNLVTGWQYRFFVLNNEAGLLEYFVNEQSRNQKPRGTLQLAGAVISPSDEDSHTFTVNAASGEQYKLRATDAKERQHWVSRLQICTQHHTEAIGKNNPPLKSRSFSLASSSNSPISQRRPSQNAISFFNVGHSKLQSLSKRTNLPPDHLVEVREMMSHAEGQQRDLIRRIECLPTSGHLSSLDQDLLMLKATSMATMNCLNDCFHILQLQHASHQKGSLPSGTTIEWLEPKISLSNHYKNGADQPFATDQSKPVAVPEEQPVAESGLLAREPEEINADDEIEDTCDHKEDDLGAVEEQRSVILHLLSQLKLGMDLTRVVLPTFILEKRSLLEMYADFMSHPDLFIAITNGATAEDRMIRFVEYYLTSFHEGRKGAIAKKPYNPIIGETFHCSWKMPKSEVASSVFSSSSTQGVTNHAPLSGESLTQVGSDCYTVRFVAEQVSHHPPVSGFYAECTERKMCVNAHVWTKSKFLGMSIGVTMVGEGILSLLEHGEEYTFSLPCAYARSILTVPWVELGGKVSVNCAKTGYSASITFHTKPFYGGKLHRVTAEVKHNITNTVVCRVQGEWNSVLEFTYSNGETKYVDLTKLAVTKKRVRPLEKQDPFESRRLWKNVTDSLRESEIDKATEHKHTLEERQRTEERHRTETGTPWKTKYFIKEGDGWVYHKPLWKIIPTTQPAE B443D3BDE8AE5FB6 pi3p 1-phosphatidyl-1D-myo-inositol 3-phosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,4-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) osb10_human Oxysterol-binding protein-related protein 10 OSBPL10 ORP10 OSBP9 protein protein human Homo sapiens Human MERAVQGTDGGGGSNSSSRSSSRATSAGSSPSCSLAGRGVSSRSAAAGLGGGGSRSSPGSVAASPSGGGGRRREPALEGVLSKYTNLLQGWQNRYFVLDFEAGILQYFVNEQSKHQKPRGVLSLSGAIVSLSDEAPHMLVVYSANGEMFKLRAADAKEKQFWVTQLRACAKYHMEMNSKSAPSSRSRSLTLLPHGTPNSASPCSQRHLSVGAPGVVTITHHKSPAAARRAKSQYSGQLHEVREMMNQVEGQQKNLVHAIESLPGSGPLTALDQDLLLLKATSAATLSCLGECLNLLQQSVHQAGQPSQKPGASENILGWHGSKSHSTEQLKNGTLGSLPSASANITWAILPNSAEDEQTSQPEPEPNSGSELVLSEDEKSDNEDKEETELGVMEDQRSIILHLISQLKLGMDLTKVVLPTFILEKRSLLEMYADFMAHPDLLLAITAGATPEERVICFVEYYLTAFHEGRKGALAKKPYNPIIGETFHCSWEVPKDRVKPKRTASRSPASCHEHPMADDPSKSYKLRFVAEQVSHHPPISCFYCECEEKRLCVNTHVWTKSKFMGMSVGVSMIGEGVLRLLEHGEEYVFTLPSAYARSILTIPWVELGGKVSINCAKTGYSATVIFHTKPFYGGKVHRVTAEVKHNPTNTIVCKAHGEWNGTLEFTYNNGETKVIDTTTLPVYPKKIRPLEKQGPMESRNLWREVTRYLRLGDIDAATEQKRHLEEKQRVEERKRENLRTPWKPKYFIQEGDGWVYFNPLWKAH 62B1DF599C7C15FC pi5p 1-phosphatidyl-1D-myo-inositol 5-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,4,5-trisphosphate 1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) phosphatidyl-l-serines phosphatidyl-L-serines small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cholesterol cholesterol small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi4p 1-phosphatidyl-1D-myo-inositol 4-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 4,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) osbpl10-pi4p-1 3 15 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A osbpl10-pi3p-1 3 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A osbpl10-1_phosphatidyl_1d_myo_inositol 3,4_bisphosphate-1 3 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A osbpl10-1_phosphatidyl_1d_myo_inositol 3,5_bisphosphate-1 3 16 unspecified role unspecified role bait bait 10 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 region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A osbpl10-1_phosphatidyl_1d_myo_inositol 4,5_bisphosphate-1 3 17 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A pi5p-osbpl10-1 3 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A 1_phosphatidyl_1d_myo_inositol 3,4,5_trisphosphate-osbpl10-1 3 12 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A phosphatidic acids-osbpl10-1 3 5 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey partially purified partially purified region 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1A phosphatidyl_l_serines-osbpl10-1 3 10 unspecified role unspecified role prey prey partially purified partially purified region 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role bait bait physical association physical association Fig. 1A cholesterol-osbpl10-1 4 14 unspecified role unspecified role neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 10 unspecified role unspecified role neutral component neutral component partially purified partially purified region 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1B Deletion of the region 412–415 (amino acid residues DLTK) resulted in an equal or even higher cholesterol extraction capacity than the wild-type OSBPL10. osbpl10-osbpl9-1 2 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n-terminus xpress tag xpress tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 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 physical association physical association Fig 5D To further confirm the association of ORP10 with ORP9L suggested by the BiFC analysis, we carried out co-immunoprecipitation of the two proteins from the lysate of HuH7 cells transfected with GFP-ORP10 either alone or together with Xpress®-epitope tagged ORP9L, followed by precipitation with the Xpress® antibody. osbpl9-osbpl10-1 1 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell region venus-c venus c-terminal fragment undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell region venus-n venus n-terminal fragment undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig 5A We therefore employed the bimolecular fluorescence complementation technique [BiFC; [37]] to visualize in live cells the interaction of ORP10 with ORP9L. osbpl11-osbpl9-5 1 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell region venus-c venus c-terminal fragment undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell region venus-n venus n-terminal fragment undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig 5B