MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Conformational changes in the AAA ATPase p97-p47 adaptor complex. Conformational changes in the AAA ATPase p97-p47 adaptor complex. EMBO J. (0261-4189) 2006 imex curation Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. 2009-01-08 Only protein-protein interactions unknown Unknown biochemical biochemical nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Conformational changes in the AAA ATPase p97-p47 adaptor complex. Conformational changes in the AAA ATPase p97-p47 adaptor complex. EMBO J. (0261-4189) 2006 imex curation Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. 2009-01-08 Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Conformational changes in the AAA ATPase p97-p47 adaptor complex. Conformational changes in the AAA ATPase p97-p47 adaptor complex. EMBO J. (0261-4189) 2006 imex curation Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. 2009-01-08 Only protein-protein interactions unknown Unknown itc isothermal titration calorimetry ITC ITC itc nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Conformational changes in the AAA ATPase p97-p47 adaptor complex. Conformational changes in the AAA ATPase p97-p47 adaptor complex. EMBO J. (0261-4189) 2006 imex curation Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. 2009-01-08 Only protein-protein interactions in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Beuron F., Dreveny I., Yuan X., Pye VE., McKeown C., Briggs LC., Cliff MJ., Kaneko Y., Wallis R., Isaacson RL., Ladbury JE., Matthews SJ., Kondo H., Zhang X., Freemont PS. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation tera_rat Transitional endoplasmic reticulum ATPase Vcp 15S Mg(2+)-ATPase p97 subunit Valosin-containing protein protein protein rat Rattus norvegicus (Rat) Rat MASGADSKGDDLSTAILKQKNRPNRLIVDEAINEDNSVVSLSQPKMDELQLFRGDTVLLKGKKRREAVCIVLSDDTCSDEKIRMNRVVRNNLRVRLGDVISIQPCPDVKYGKRIHVLPIDDTVEGITGNLFEVYLKPYFLEAYRPIRKGDIFLVRGGMRAVEFKVVETDPSPYCIVAPDTVIHCEGEPIKREDEEESLNEVGYDDIGGCRKQLAQIKEMVELPLRHPALFKAIGVKPPRGILLYGPPGTGKTLIARAVANETGAFFFLINGPEIMSKLAGESESNLRKAFEEAEKNAPAIIFIDELDAIAPKREKTHGEVERRIVSQLLTLMDGLKQRAHVIVMAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEILQIHTKNMKLADDVDLEQVANETHGHVGADLAALCSEAALQAIRKKMDLIDLEDETIDAEVMNSLAVTMDDFRWALSQSNPSALRETVVEVPQVTWEDIGGLEDVKRELQELVQYPVEHPDKFLKFGMTPSKGVLFYGPPGCGKTLLAKAIANECQANFISIKGPELLTMWFGESEANVREIFDKARQAAPCVLFFDELDSIAKARGGNIGDGGGAADRVINQILTEMDGMSTKKNVFIIGATNRPDIIDPAILRPGRLDQLIYIPLPDEKSRVAILKANLRKSPVAKDVDLEFLAKMTNGFSGADLTEICQRACKLAIRESIESEIRRERERQTNPSAMEVEEDDPVPEIRRDHFEEAMRFARRSVSDNDIRKYEMFAQTLQQSRGFGSFRFPSGNQGGAGPSQGSGGGTGGNVYTEDNDDDLYG 501B721D205EBA8A nsf1c_rat NSFL1 cofactor p47 Nsfl1c p97 cofactor p47 XY body-associated protein XY40 protein protein rat Rattus norvegicus (Rat) Rat MAEERQDALREFVAVTGAEEDRARFFLESAGWDLQIALASFYEDGGDEDIVTISQATPSSVSRGTAPSDNRVTSFRDLIHDQDEEEEEEEGQRFYAGGSERSGQQIVGPPRKKSPNELVDDLFKGAKEHGAVAVERVTKSPGETSKPRPFAGGGYRLGAAPEEESAYVAGERRRHSGQDVHVVLKLWKTGFSLDNGDLRSYQDPSNAQFLESIRRGEVPAELRRLAHGGQVNLDMEDHRDEDFVKPKGAFKAFTGEGQKLGSTAPQVLNTSSPAQQAENEAKASSSILINEAEPTTNIQIRLADGGRLVQKFNHSHRISDIRLFIVDARPAMAATSFVLMTTFPNKELADENQTLKEANLLNAVIVQRLT 60C81402E5C58134 nsfl1c-nsfl1c 1 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged 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 Stoichiometry: 1.0 direct interaction direct interaction EMBO-Journal SF1B p47 existis as a trimer Furthermore, analytical ultracentrifugation data presented in this study provides additional evidence for full-length p47 existing as a trimer (Supplementary Figure 1B). kd 0.11 nanomolar homomint mint nsfl1c-vcp-6 3 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged 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 Stoichiometry: 1.0 direct interaction direct interaction F1C EMBO-Journal A ratio of 0.5 p47 monomer to 1 p97 protomer was observed confirming that three p47 bind to a hexameric p97 A ratio of 0.5 p47 monomer to 1 p97 protomer was observed with a Kd of 0.5 mM confirming that three p47 bind to a hexameric p97 (Figure 1C) kd 0.5 micromolar homomint mint nsfl1c-vcp-7 3 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-1954443: original sequence: 171-370 : ERRRHSGQDVHVVLKLWKTGFSLDNGDLRSYQDPSNAQFLESIRRGEVPAELRRLAHGGQVNLDMEDHRDEDFVKPKGAFKAFTGEGQKLGSTAPQVLNTSSPAQQAENEAKASSSILINEAEPTTNIQIRLADGGRLVQKFNHSHRISDIRLFIVDARPAMAATSFVLMTTFPNKELADENQTLKEANLLNAVIVQRL Stoichiometry: 1.0 direct interaction direct interaction F1D EMBO-Journal kd 0.5 micromolar Ihe stoichiometry of a shorter p47 fragment, comprising the SEP UBX domains, was found to be of 1 p47 fragment bound to 1 p97 protomer In contrast, the stoichiometry of a shorter p47 fragment, comprising the SEP UBX domains, was found to be of 1 p47 fragment bound to 1 p97 protomer homomint mint nsfl1c-vcp-8 4 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 direct interaction direct interaction F3 F2 EMBO-Journal We have obtained cryomicrographs of p97 p47 complex in the presence of ADP or AMPPNP (Figure 2 and Supplementary Table I). Manual picking of single particles was straightforward because of the good contrast of the particles (Figure 2A) homomint mint nsfl1c-vcp-9 2 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 colocalization colocalization SF2 EMBO-Journal Scanning transmission electron microscopy mass measurements also confirm that the particles are no larger than a p97 hexamer in complex with 3 p47 (data not shown) consistent with the calorimetric data and analysis of the complex by native gel electrophoresis (Figure 1C and Supplementary Figure 2A). homomint mint