UniProt UniProt http://www.uniprot.org kondo-1997-1 p47 is a cofactor for p97-mediated membrane fusion. rat-liver rattus norvegicus liver Rat liver liver molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Kondo H., Rabouille C., Newman R., Levine TP., Pappin D., Freemont P., Warren G. Nsfl1c/p47 was also partial sequenced by mass spectrometry (extracted from gel slices). g.warren@europa.lif.icnet.uk Nature (0028-0836) 1997 Only protein-protein interactions imex curation kondo-1997-2 p47 is a cofactor for p97-mediated membrane fusion. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight by comassie molecular weight estimation by coomasie staining weight by comassie Kondo H., Rabouille C., Newman R., Levine TP., Pappin D., Freemont P., Warren G. Nsfl1c/p47 was also partial sequenced by mass spectrometry (extracted from gel slices). Crude cytosol was added to recombinant His6-p47/Nsfl1c followed by antibodies against the His6 tag. The immunoprecipitate was then fractionated by SDS_PAGE and the gel stained with Coomassie blue. g.warren@europa.lif.icnet.uk Nature (0028-0836) 1997 Only protein-protein interactions imex curation kondo-1997-3 p47 is a cofactor for p97-mediated membrane fusion. rat-liver rattus norvegicus liver Rat liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip antibody detection identification by antibody antibody detection Kondo H., Rabouille C., Newman R., Levine TP., Pappin D., Freemont P., Warren G. Nsfl1c/p47 was also partial sequenced by mass spectrometry (extracted from gel slices). g.warren@europa.lif.icnet.uk Nature (0028-0836) 1997 Only protein-protein interactions imex curation kondo-1997-4 p47 is a cofactor for p97-mediated membrane fusion. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Kondo H., Rabouille C., Newman R., Levine TP., Pappin D., Freemont P., Warren G. Nsfl1c/p47 was also partial sequenced by mass spectrometry (extracted from gel slices). The reconstituted complex was prepared by mixing pure p97 with excess His6-p47, isolated by gel filtration. Fractions assayed by Western blotting, also visualized by Coomassie staining. g.warren@europa.lif.icnet.uk Nature (0028-0836) 1997 Only protein-protein interactions imex curation nsf1c_rat NSFL1 cofactor p47 Nsfl1c p97 cofactor p47 XY body-associated protein XY40 protein protein rat Rattus norvegicus (Rat) Rat MAEERQDALREFVAVTGAEEDRARFFLESAGWDLQIALASFYEDGGDEDIVTISQATPSSVSRGTAPSDNRVTSFRDLIHDQDEEEEEEEGQRFYAGGSERSGQQIVGPPRKKSPNELVDDLFKGAKEHGAVAVERVTKSPGETSKPRPFAGGGYRLGAAPEEESAYVAGERRRHSGQDVHVVLKLWKTGFSLDNGDLRSYQDPSNAQFLESIRRGEVPAELRRLAHGGQVNLDMEDHRDEDFVKPKGAFKAFTGEGQKLGSTAPQVLNTSSPAQQAENEAKASSSILINEAEPTTNIQIRLADGGRLVQKFNHSHRISDIRLFIVDARPAMAATSFVLMTTFPNKELADENQTLKEANLLNAVIVQRLT tera_rat Transitional endoplasmic reticulum ATPase Vcp 15S Mg(2+)-ATPase p97 subunit Valosin-containing protein protein protein rat Rattus norvegicus (Rat) Rat MASGADSKGDDLSTAILKQKNRPNRLIVDEAINEDNSVVSLSQPKMDELQLFRGDTVLLKGKKRREAVCIVLSDDTCSDEKIRMNRVVRNNLRVRLGDVISIQPCPDVKYGKRIHVLPIDDTVEGITGNLFEVYLKPYFLEAYRPIRKGDIFLVRGGMRAVEFKVVETDPSPYCIVAPDTVIHCEGEPIKREDEEESLNEVGYDDIGGCRKQLAQIKEMVELPLRHPALFKAIGVKPPRGILLYGPPGTGKTLIARAVANETGAFFFLINGPEIMSKLAGESESNLRKAFEEAEKNAPAIIFIDELDAIAPKREKTHGEVERRIVSQLLTLMDGLKQRAHVIVMAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEILQIHTKNMKLADDVDLEQVANETHGHVGADLAALCSEAALQAIRKKMDLIDLEDETIDAEVMNSLAVTMDDFRWALSQSNPSALRETVVEVPQVTWEDIGGLEDVKRELQELVQYPVEHPDKFLKFGMTPSKGVLFYGPPGCGKTLLAKAIANECQANFISIKGPELLTMWFGESEANVREIFDKARQAAPCVLFFDELDSIAKARGGNIGDGGGAADRVINQILTEMDGMSTKKNVFIIGATNRPDIIDPAILRPGRLDQLIYIPLPDEKSRVAILKANLRKSPVAKDVDLEFLAKMTNGFSGADLTEICQRACKLAIRESIESEIRRERERQTNPSAMEVEEDDPVPEIRRDHFEEAMRFARRSVSDNDIRKYEMFAQTLQQSRGFGSFRFPSGNQGGAGPSQGSGGGTGGNVYTEDNDDDLYG vcp-nsfl1c-3 777128 n/a 777132 unspecified role unspecified role neutral component neutral component n/a 777133 unspecified role unspecified role neutral component neutral component physical association physical association false false false 2a vcp-nsfl1c-4 777129 n/a 777133 unspecified role unspecified role prey prey n/a 777132 unspecified role unspecified role bait bait n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false in vitro In vitro physical association physical association false false false 2c nsfl1c-vcp 777130 n/a 777132 unspecified role unspecified role bait bait n/a 777133 unspecified role unspecified role prey prey physical association physical association false false false 2b vcp-nsfl1c-5 777131 n/a 777133 unspecified role unspecified role neutral component neutral component n/a 777132 unspecified role unspecified role neutral component neutral component n-terninus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false in vitro In vitro physical association physical association false false false 1c and 2a