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/ Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard in vitro In vitro peptide array peptide array predetermined predetermined participant predetermined Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. The origin of the UBCH5B is determined from the vendors site and the origins of the enzyme E1 determined from the back reference PMID: 11676916 and the vendors site. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Identification of VCP/p97, carboxyl terminus of Hsp70-interacting protein (CHIP), and amphiphysin II interaction partners using membrane-based human proteome arrays. Mol. Cell Proteomics (1535-9476) 2006 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Synapse - Interactions of proteins with an established role in the presynapse. Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. orchard 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 Grelle G., Kostka S., Otto A., Kersten B., Genser KF., Muller EC., Walter S., Boddrich A., Stelzl U., Hanig C., Volkmer-Engert R., Landgraf C., Alberti S., Hohfeld J., Strodicke M., Wanker EE. Parkinsons - Interactions investigated in the context of Parkinsons disease Crude bacterial protein extracts containing His-tagged human fusion proteins are double spotted onto filter membranes. PPIs are detected after overlay with bacterial protein extracts containing recombinant human GST fusion protein using HRP-conjugated anti-GST antibody. The GST tagged bait determined by ELISA and the HIS tagged preys determined by mass spectrometry. ewanker@mdc-berlin.de Accepted 2008-NOV-13 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Human fetal brain cDNA library were used for His-tagged fusion proteins. Mol. Cell Proteomics (1535-9476) 2006 orchard dlgp4_human Disks large-associated protein 4 SAPAP4 SAP90/PSD-95-associated protein 4 PSD-95/SAP90-binding protein 4 DLGAP4 DAP4 KIAA0964 protein protein human Homo sapiens Human MKGLGDSRPRHLSDSLDPPHEPLFAGTDRNPYLLSPTEAFAREARFPGQNTLPGDGLFPLNNQLPPPSSTFPRIHYNSHFEVPEESPFPSHAQATKINRLPANLLDQFEKQLPIHRDGFSTLQFPRGEAKARGESPGRIRHLVHSVQRLFFTKAPSLEGTAGKVGGNGSKKGGMEDGKGRRAKSKERAKAGEPKRRSRSNISGWWSSDDNLDGEAGAFRSSGPASGLMTLGRQAERSQPRYFMHAYNTISGHMLKTTKNNTTELTAPPPPPAPPATCPSLGVGTDTNYVKRGSWSTLTLSHAHEVCQKTSATLDKSLLKSKSCHQGLAYHYLQVPGGGGEWSTTLLSPRETDAAAEGPIPCRRMRSGSYIKAMGDEDSDESGGSPKPSPKTAARRQSYLRATQQSLGEQSNPRRSLDRLDSVDMLLPSKCPSWEEDYTPVSDSLNDSSCISQIFGQASLIPQLFGHEQQVREAELSDQYEAACESACSEAESTAAETLDLPLPSYFRSRSHSYLRAIQAGCSQEEDSVSLQSLSPPPSTGSLSNSRTLPSSSCLVAYKKTPPPVPPRTTSKPFISVTVQSSTESAQDTYLDSQDHKSEVTSQSGLSNSSDSLDSSTRPPSVTRGGVAPAPEAPEPPPKHAALKSEQGTLTSSESHPEAAPKRKLSSIGIQVDCIQPVPKEEPSPATKFQSIGVQVEDDWRSSVPSHSMSSRRDTDSDTQDANDSSCKSSERSLPDCTPHPNSISIDAGPRQAPKIAQIKRNLSYGDNSDPALEASSLPPPDPWLETSSSSPAEPAQPGACRRDGYWFLKLLQAETERLEGWCCQMDKETKENNLSEEVLGKVLSAVGSAQLLMSQKFQQFRGLCEQNLNPDANPRPTAQDLAGFWDLLQLSIEDISMKFDELYHLKANSWQLVETPEKRKEEKKPPPPVPKKPAKSKPAVSRDKASDASDKQRQEARKRLLAAKRAASVRQNSATESADSIEIYVPEAQTRL F0EBFBE0DE1AC814 nsf1c_human NSFL1 cofactor p47 p97 cofactor p47 NSFL1C UBXN2C UBX domain-containing protein 2C protein protein human Homo sapiens Human MAAERQEALREFVAVTGAEEDRARFFLESAGWDLQIALASFYEDGGDEDIVTISQATPSSVSRGTAPSDNRVTSFRDLIHDQDEDEEEEEGQRFYAGGSERSGQQIVGPPRKKSPNELVDDLFKGAKEHGAVAVERVTKSPGETSKPRPFAGGGYRLGAAPEEESAYVAGEKRQHSSQDVHVVLKLWKSGFSLDNGELRSYQDPSNAQFLESIRRGEVPAELRRLAHGGQVNLDMEDHRDEDFVKPKGAFKAFTGEGQKLGSTAPQVLSTSSPAQQAENEAKASSSILIDESEPTTNIQIRLADGGRLVQKFNHSHRISDIRLFIVDARPAMAATSFILMTTFPNKELADESQTLKEANLLNAVIVQRLT DIP protein Q9UNZ2 original sequence version: 104 DIP protein Q9UNZ2 original sequence version: 145 DIP protein Q9UNZ2 original sequence version: 112 79364617F940B9F9 hsp7c_human Heat shock cognate 71 kDa protein HSPA8 HSC70 HSP73 HSPA10 Heat shock 70 kDa protein 8 Lipopolysaccharide-associated protein 1 protein protein human Homo sapiens Human MSKGPAVGIDLGTTYSCVGVFQHGKVEIIANDQGNRTTPSYVAFTDTERLIGDAAKNQVAMNPTNTVFDAKRLIGRRFDDAVVQSDMKHWPFMVVNDAGRPKVQVEYKGETKSFYPEEVSSMVLTKMKEIAEAYLGKTVTNAVVTVPAYFNDSQRQATKDAGTIAGLNVLRIINEPTAAAIAYGLDKKVGAERNVLIFDLGGGTFDVSILTIEDGIFEVKSTAGDTHLGGEDFDNRMVNHFIAEFKRKHKKDISENKRAVRRLRTACERAKRTLSSSTQASIEIDSLYEGIDFYTSITRARFEELNADLFRGTLDPVEKALRDAKLDKSQIHDIVLVGGSTRIPKIQKLLQDFFNGKELNKSINPDEAVAYGAAVQAAILSGDKSENVQDLLLLDVTPLSLGIETAGGVMTVLIKRNTTIPTKQTQTFTTYSDNQPGVLIQVYEGERAMTKDNNLLGKFELTGIPPAPRGVPQIEVTFDIDANGILNVSAVDKSTGKENKITITNDKGRLSKEDIERMVQEAEKYKAEDEKQRDKVSSKNSLESYAFNMKATVEDEKLQGKINDEDKQKILDKCNEIINWLDKNQTAEKEEFEHQQKELEKVCNPIITKLYQSAGGMPGGMPGGFPGGGAPPSGGASSGPTIEEVD 9AA27B210730670C atcay_human Caytaxin ATCAY KIAA1872 Ataxia cayman type protein BNIP-2-homology protein protein human Homo sapiens Human MGTTEATLRMENVDVKEEWQDEDLPRPLPEETGVELLGSPVEDTSSPPNTLNFNGAHRKRKTLVAPEINISLDQSEGSLLSDDFLDTPDDLDINVDDIETPDETDSLEFLGNGNELEWEDDTPVATAKNMPGDSADLFGDGTTEDGSAANGRLWRTVIIGEQEHRIDLHMIRPYMKVVTHGGYYGEGLNAIIVFAACFLPDSSLPDYHYIMENLFLYVISSLELLVAEDYMIVYLNGATPRRRMPGIGWLKKCYQMIDRRLRKNLKSLIIVHPSWFIRTVLAISRPFISVKFINKIQYVHSLEDLEQLIPMEHVQIPDCVLQYEEERLKARRESARPQPEFVLPRSEEKPEVAPVENRSALVSEDQETSMS C09174CE3AD77764 dyn1_human Dynamin-1 DNM1 DNM protein protein human Homo sapiens Human MGNRGMEDLIPLVNRLQDAFSAIGQNADLDLPQIAVVGGQSAGKSSVLENFVGRDFLPRGSGIVTRRPLVLQLVNATTEYAEFLHCKGKKFTDFEEVRLEIEAETDRVTGTNKGISPVPINLRVYSPHVLNLTLVDLPGMTKVPVGDQPPDIEFQIRDMLMQFVTKENCLILAVSPANSDLANSDALKVAKEVDPQGQRTIGVITKLDLMDEGTDARDVLENKLLPLRRGYIGVVNRSQKDIDGKKDITAALAAERKFFLSHPSYRHLADRMGTPYLQKVLNQQLTNHIRDTLPGLRNKLQSQLLSIEKEVEEYKNFRPDDPARKTKALLQMVQQFAVDFEKRIEGSGDQIDTYELSGGARINRIFHERFPFELVKMEFDEKELRREISYAIKNIHGIRTGLFTPDMAFETIVKKQVKKIREPCLKCVDMVISELISTVRQCTKKLQQYPRLREEMERIVTTHIREREGRTKEQVMLLIDIELAYMNTNHEDFIGFANAQQRSNQMNKKKTSGNQDEILVIRKGWLTINNIGIMKGGSKEYWFVLTAENLSWYKDDEEKEKKYMLSVDNLKLRDVEKGFMSSKHIFALFNTEQRNVYKDYRQLELACETQEEVDSWKASFLRAGVYPERVGDKEKASETEENGSDSFMHSMDPQLERQVETIRNLVDSYMAIVNKTVRDLMPKTIMHLMINNTKEFIFSELLANLYSCGDQNTLMEESAEQAQRRDEMLRMYHALKEALSIIGDINTTTVSTPMPPPVDDSWLQVQSVPAGRRSPTSSPTPQRRAPAVPPARPGSRGPAPGPPPAGSALGGAPPVPSRPGASPDPFGPPPQVPSRPNRAPPGVPSRSGQASPSRPESPRPPFDL DIP protein Q05193 original sequence version: 142 DIP protein Q05193 original sequence version: 113 DIP protein Q05193 original sequence version: 100 DIP protein Q05193 original sequence version: 115 DIP protein Q05193-1 original sequence version: 111 DIP protein Q05193 original sequence version: 153 7FCD8CB572FFEAEF amfr_human E3 ubiquitin-protein ligase AMFR gp78 RING finger protein 45 AMFR RNF45 Autocrine motility factor receptor protein protein human Homo sapiens Human MPLLFLERFPWPSLRTYTGLSGLALLGTIISAYRALSQPEAGPGEPDQLTASLQPEPPAPARPSAGGPRARDVAQYLLSDSLFVWVLVNTACCVLMLVAKLIQCIVFGPLRVSERQHLKDKFWNFIFYKFIFIFGVLNVQTVEEVVMWCLWFAGLVFLHLMVQLCKDRFEYLSFSPTTPMSSHGRVLSLLVAMLLSCCGLAAVCSITGYTHGMHTLAFMAAESLLVTVRTAHVILRYVIHLWDLNHEGTWEGKGTYVYYTDFVMELTLLSLDLMHHIHMLLFGNIWLSMASLVIFMQLRYLFHEVQRRIRRHKNYLRVVGNMEARFAVATPEELAVNNDDCAICWDSMQAARKLPCGHLFHNSCLRSWLEQDTSCPTCRMSLNIADNNRVREEHQGENLDENLVPVAAAEGRPRLNQHNHFFHFDGSRIASWLPSFSVEVMHTTNILGITQASNSQLNAMAHQIQEMFPQVPYHLVLQDLQLTRSVEITTDNILEGRIQVPFPTQRSDSIRPALNSPVERPSSDQEEGETSAQTERVPLDLSPRLEETLDFGEVEVEPSEVEDFEARGSRFSKSADERQRMLVQRKDELLQQARKRFLNKSSEDDAASESFLPSEGASSDPVTLRRRMLAAAAERRLQKQQTS DIP protein Q9UKV5 original sequence version: 136 DIP protein Q9UKV5 original sequence version: 122 DIP protein Q9UKV5 original sequence version: 113 DIP protein Q9UKV5 original sequence version: 85 DIP protein Q9UKV5 original sequence version: 103 DIP protein Q9UKV5 original sequence version: 40 8782324609C0E62A bin1_human Myc box-dependent-interacting protein 1 Bridging integrator 1 Amphiphysin-like protein Box-dependent myc-interacting protein 1 BIN1 AMPHL Amphiphysin II protein protein human Homo sapiens Human MAEMGSKGVTAGKIASNVQKKLTRAQEKVLQKLGKADETKDEQFEQCVQNFNKQLTEGTRLQKDLRTYLASVKAMHEASKKLNECLQEVYEPDWPGRDEANKIAENNDLLWMDYHQKLVDQALLTMDTYLGQFPDIKSRIAKRGRKLVDYDSARHHYESLQTAKKKDEAKIAKPVSLLEKAAPQWCQGKLQAHLVAQTNLLRNQAEEELIKAQKVFEEMNVDLQEELPSLWNSRVGFYVNTFQSIAGLEENFHKEMSKLNQNLNDVLVGLEKQHGSNTFTVKAQPSDNAPAKGNKSPSPPDGSPAATPEIRVNHEPEPAGGATPGATLPKSPSQLRKGPPVPPPPKHTPSKEVKQEQILSLFEDTFVPEISVTTPSQFEAPGPFSEQASLLDLDFDPLPPVTSPVKAPTPSGQSIPWDLWEPTESPAGSLPSGEPSAAEGTFAVSWPSQTAEPGPAQPAEASEVAGGTQPAAGAQEPGETAASEAASSSLPAVVVETFPATVNGTVEGGSGAGRLDLPPGFMFKVQAQHDYTATDTDELQLKAGDVVLVIPFQNPEEQDEGWLMGVKESDWNQHKELEKCRGVFPENFTERVP 0FF1956F0C7E3B50 ub2d2_human Ubiquitin-conjugating enzyme E2 D2 UBC5B UBCH4 UBE2D2 UBC4 UBCH5B Ubiquitin-protein ligase D2 Ubiquitin carrier protein D2 Ubiquitin-conjugating enzyme E2-17 kDa 2 Ubiquitin-conjugating enzyme E2(17)KB 2 PUBC1 p53-regulated ubiquitin-conjugating enzyme 1 (E3-independent) E2 ubiquitin-conjugating enzyme D2 E2 ubiquitin-conjugating enzyme D2 protein protein human Homo sapiens Human MALKRIHKELNDLARDPPAQCSAGPVGDDMFHWQATIMGPNDSPYQGGVFFLTIHFPTDYPFKPPKVAFTTRIYHPNINSNGSICLDILRSQWSPALTISKVLLSICSLLCDPNPDDPLVPEIARIYKTDREKYNRIAREWTQKYAM DIP protein P62837 original sequence version: 81 DIP protein P62837-1 original sequence version: 134 DIP protein P62837 original sequence version: 95 DIP protein P62837 original sequence version: 139 DIP protein P62837 original sequence version: 37 DIP protein P62837 original sequence version: 115 DIP protein P62837 original sequence version: 58 DIP protein P62837 original sequence version: 74 DIP protein P62837 original sequence version: 103 DIP protein P62837 original sequence version: 101 C942BE7853CBC355 xrcc4_human DNA repair protein XRCC4 X-ray repair cross-complementing protein 4 XRCC4 protein protein human Homo sapiens Human MERKISRIHLVSEPSITHFLQVSWEKTLESGFVITLTDGHSAWTGTVSESEISQEADDMAMEKGKYVGELRKALLSGAGPADVYTFNFSKESCYFFFEKNLKDVSFRLGSFNLEKVENPAEVIRELICYCLDTIAENQAKNEHLQKENERLLRDWNDVQGRFEKCVSAKEALETDLYKRFILVLNEKKTKIRSLHNKLLNAAQEREKDIKQEGETAICSEMTADRDPVYDESTDEESENQTDLSGLASAAVSKDDSIISSLDVTDIAPSRKRRQRMQRNLGTEPKMAPQENQLQEKENSRPDSSLPETSKKEHISAENMSLETLRNSSPEDLFDEI DIP protein Q13426 original sequence version: 140 DIP protein Q13426-1 original sequence version: 116 BE5FB99153479A4E tera_human Transitional endoplasmic reticulum ATPase VCP 15S Mg(2+)-ATPase p97 subunit Valosin-containing protein protein protein human Homo sapiens Human MASGADSKGDDLSTAILKQKNRPNRLIVDEAINEDNSVVSLSQPKMDELQLFRGDTVLLKGKKRREAVCIVLSDDTCSDEKIRMNRVVRNNLRVRLGDVISIQPCPDVKYGKRIHVLPIDDTVEGITGNLFEVYLKPYFLEAYRPIRKGDIFLVRGGMRAVEFKVVETDPSPYCIVAPDTVIHCEGEPIKREDEEESLNEVGYDDIGGCRKQLAQIKEMVELPLRHPALFKAIGVKPPRGILLYGPPGTGKTLIARAVANETGAFFFLINGPEIMSKLAGESESNLRKAFEEAEKNAPAIIFIDELDAIAPKREKTHGEVERRIVSQLLTLMDGLKQRAHVIVMAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEILQIHTKNMKLADDVDLEQVANETHGHVGADLAALCSEAALQAIRKKMDLIDLEDETIDAEVMNSLAVTMDDFRWALSQSNPSALRETVVEVPQVTWEDIGGLEDVKRELQELVQYPVEHPDKFLKFGMTPSKGVLFYGPPGCGKTLLAKAIANECQANFISIKGPELLTMWFGESEANVREIFDKARQAAPCVLFFDELDSIAKARGGNIGDGGGAADRVINQILTEMDGMSTKKNVFIIGATNRPDIIDPAILRPGRLDQLIYIPLPDEKSRVAILKANLRKSPVAKDVDLEFLAKMTNGFSGADLTEICQRACKLAIRESIESEIRRERERQTNPSAMEVEEDDPVPEIRRDHFEEAMRFARRSVSDNDIRKYEMFAQTLQQSRGFGSFRFPSGNQGGAGPSQGSGGGTGGSVYTEDNDDDLYG DIP protein P55072 original sequence version: 89 DIP protein P55072 original sequence version: 114 DIP protein P55072 original sequence version: 123 DIP protein P55072 original sequence version: 150 DIP protein P55072 original sequence version: 174 DIP protein P55072 original sequence version: 175 DIP protein P55072 original sequence version: 176 DIP protein P55072 original sequence version: 160 DIP protein P55072 original sequence version: 128 DIP protein P55072 original sequence version: 132 501B721D3A77BA8A tau_human Microtubule-associated protein tau MAPT MTBT1 TAU Neurofibrillary tangle protein Paired helical filament-tau MAPTL protein protein human Homo sapiens Human MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQEPESGKVVQEGFLREPGPPGLSHQLMSGMPGAPLLPEGPREATRQPSGTGPEDTEGGRHAPELLKHQLLGDLHQEGPPLKGAGGKERPGSKEEVDEDRDVDESSPQDSPPSKASPAQDGRPPQTAAREATSIPGFPAEGAIPLPVDFLSKVSTEIPASEPDGPSVGRAKGQDAPLEFTFHVEITPNVQKEQAHSEEHLGRAAFPGAPGEGPEARGPSLGEDTKEADLPEPSEKQPAAAPRGKPVSRVPQLKARMVSKSKDGTGSDDKKAKTSTRSSAKTLKNRPCLSPKHPTPGSSDPLIQPSSPAVCPEPPSSPKYVSSVTSRTGSSGAKEMKLKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL DIP protein P10636 original sequence version: 125 DIP protein P10636 original sequence version: 210 D46C66CDBCD196E8 hsp71_human Heat shock 70 kDa protein 1A/1B HSPA1A HSPA1 HSPA1B Heat shock 70 kDa protein 1/2 HSX70 protein protein human Homo sapiens Human MAKAAAIGIDLGTTYSCVGVFQHGKVEIIANDQGNRTTPSYVAFTDTERLIGDAAKNQVALNPQNTVFDAKRLIGRKFGDPVVQSDMKHWPFQVINDGDKPKVQVSYKGETKAFYPEEISSMVLTKMKEIAEAYLGYPVTNAVITVPAYFNDSQRQATKDAGVIAGLNVLRIINEPTAAAIAYGLDRTGKGERNVLIFDLGGGTFDVSILTIDDGIFEVKATAGDTHLGGEDFDNRLVNHFVEEFKRKHKKDISQNKRAVRRLRTACERAKRTLSSSTQASLEIDSLFEGIDFYTSITRARFEELCSDLFRSTLEPVEKALRDAKLDKAQIHDLVLVGGSTRIPKVQKLLQDFFNGRDLNKSINPDEAVAYGAAVQAAILMGDKSENVQDLLLLDVAPLSLGLETAGGVMTALIKRNSTIPTKQTQIFTTYSDNQPGVLIQVYEGERAMTKDNNLLGRFELSGIPPAPRGVPQIEVTFDIDANGILNVTATDKSTGKANKITITNDKGRLSKEEIERMVQEAEKYKAEDEVQRERVSAKNALESYAFNMKSAVEDEGLKGKISEADKKKVLDKCQEVISWLDANTLAEKDEFEHKRKELEQVCNPIISGLYQGAGGPGPGGFGAQGPKGGSGSGPTIEEVD 78F513118C96DE66 hs90a_human Heat shock protein HSP 90-alpha HSP90AA1 HSP90A HSPC1 Heat shock 86 kDa Renal carcinoma antigen NY-REN-38 HSPCA Lipopolysaccharide-associated protein 2 protein protein human Homo sapiens Human MPEETQTQDQPMEEEEVETFAFQAEIAQLMSLIINTFYSNKEIFLRELISNSSDALDKIRYESLTDPSKLDSGKELHINLIPNKQDRTLTIVDTGIGMTKADLINNLGTIAKSGTKAFMEALQAGADISMIGQFGVGFYSAYLVAEKVTVITKHNDDEQYAWESSAGGSFTVRTDTGEPMGRGTKVILHLKEDQTEYLEERRIKEIVKKHSQFIGYPITLFVEKERDKEVSDDEAEEKEDKEEEKEKEEKESEDKPEIEDVGSDEEEEKKDGDKKKKKKIKEKYIDQEELNKTKPIWTRNPDDITNEEYGEFYKSLTNDWEDHLAVKHFSVEGQLEFRALLFVPRRAPFDLFENRKKKNNIKLYVRRVFIMDNCEELIPEYLNFIRGVVDSEDLPLNISREMLQQSKILKVIRKNLVKKCLELFTELAEDKENYKKFYEQFSKNIKLGIHEDSQNRKKLSELLRYYTSASGDEMVSLKDYCTRMKENQKHIYYITGETKDQVANSAFVERLRKHGLEVIYMIEPIDEYCVQQLKEFEGKTLVSVTKEGLELPEDEEEKKKQEEKKTKFENLCKIMKDILEKKVEKVVVSNRLVTSPCCIVTSTYGWTANMERIMKAQALRDNSTMGYMAAKKHLEINPDHSIIETLRQKAEADKNDKSVKDLVILLYETALLSSGFSLEDPQTHANRIYRMIKLGLGIDEDDPTADDTSAAVTEEMPPLEGDDDTSRMEEVD DIP protein P07900 original sequence version: 197 DIP protein P07900 original sequence version: 116 DIP protein P07900 original sequence version: 150 DIP protein P07900-1 original sequence version: 158 DIP protein P07900 original sequence version: 207 DIP protein P07900 original sequence version: 192 DIP protein P07900 original sequence version: 156 969F65FCC0BC86FD ub2q1_human Ubiquitin-conjugating enzyme E2 Q1 UBE2Q1 NICE5 UBE2Q PRO3094 Ubiquitin-protein ligase Q1 Ubiquitin carrier protein Q1 Protein NICE-5 E2 ubiquitin-conjugating enzyme Q1 protein protein human Homo sapiens Human MQQPQPQGQQQPGPGQQLGGQGAAPGAGGGPGGGPGPGPCLRRELKLLESIFHRGHERFRIASACLDELSCEFLLAGAGGAGAGAAPGPHLPPRGSVPGDPVRIHCNITESYPAVPPIWSVESDDPNLAAVLERLVDIKKGNTLLLQHLKRIISDLCKLYNLPQHPDVEMLDQPLPAEQCTQEDVSSEDEDEEMPEDTEDLDHYEMKEEEPAEGKKSEDDGIGKENLAILEKIKKNQRQDYLNGAVSGSVQATDRLMKELRDIYRSQSFKGGNYAVELVNDSLYDWNVKLLKVDQDSALHNDLQILKEKEGADFILLNFSFKDNFPFDPPFVRVVSPVLSGGYVLGGGAICMELLTKQGWSSAYSIESVIMQISATLVKGKARVQFGANKSQYSLTRAQQSYKSLVQIHEKNGWYTPPKEDG 106FF7E59DF65555 f16p1_human Fructose-1,6-bisphosphatase 1 D-fructose-1,6-bisphosphate 1-phosphohydrolase 1 FBP1 FBP Liver FBPase protein protein human Homo sapiens Human MADQAPFDTDVNTLTRFVMEEGRKARGTGELTQLLNSLCTAVKAISSAVRKAGIAHLYGIAGSTNVTGDQVKKLDVLSNDLVMNMLKSSFATCVLVSEEDKHAIIVEPEKRGKYVVCFDPLDGSSNIDCLVSVGTIFGIYRKKSTDEPSEKDALQPGRNLVAAGYALYGSATMLVLAMDCGVNCFMLDPAIGEFILVDKDVKIKKKGKIYSLNEGYARDFDPAVTEYIQRKKFPPDNSAPYGARYVGSMVADVHRTLVYGGIFLYPANKKSPNGKLRLLYECNPMAYVMEKAGGMATTGKEAVLDVIPTDIHQRAPVILGSPDDVLEFLKVYEKHSAQ DIP protein P09467 original sequence version: 162 B3D270BCBB358B71 uba1_rabit Ubiquitin-like modifier-activating enzyme 1 Ubiquitin-activating enzyme E1 UBA1 UBE1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MSSSPLSKKRRVSGPDPKPGSNCSPAQSVLPQVPSAPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNSPLEDQLRVGEFCHSRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSTMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMTELNGNQPIEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLSASLAEPDFVMTDFAKFSRPAQLHIGFQALHKFCAQHSRPPRPRNEEDAAELVTLARAVNSKASSAVQQDSLDEDLIRNLAFVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKESLTEDKCLPRQNRYDGQVAVFGSDLQEKLGRQKYFLVGAGAIGCELLKNFAMIGLGCGENGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVHQMNPHIRVTSHQNRVGPDTERIYDDDFFQTLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQLPQSWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVSNPLHLDYVMAAANLFAQTYGLAGSQDRAAVATLLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPPADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRHLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR 1196EAC5EEEC06A5 chip_human E3 ubiquitin-protein ligase CHIP CHIP STUB1 PP1131 Carboxy terminus of Hsp70-interacting protein STIP1 homology and U box-containing protein 1 CLL-associated antigen KW-8 Antigen NY-CO-7 RING-type E3 ubiquitin transferase CHIP protein protein human Homo sapiens Human MKGKEEKEGGARLGAGGGSPEKSPSAQELKEQGNRLFVGRKYPEAAACYGRAITRNPLVAVYYTNRALCYLKMQQHEQALADCRRALELDGQSVKAHFFLGQCQLEMESYDEAIANLQRAYSLAKEQRLNFGDDIPSALRIAKKKRWNSIEERRIHQESELHSYLSRLIAAERERELEECQRNHEGDEDDSHVRAQQACIEAKHDKYMADMDELFSQVDEKRKKRDIPDYLCGKISFELMREPCITPSGITYDRKDIEEHLQRVGHFDPVTRSPLTQEQLIPNLAMKEVIDAFISENGWVEDY 7E7D6568B17070BF vcp-nsfl1 7 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 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 ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role bait bait 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 32 28 physical association physical association Table 1 and Supplemental figure 1 Overlay experiments with VCP (Supplemental Fig. 1) led to the identification of the well known interaction partner p47 vcp-amfr 7 14 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role bait bait 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 36 39 physical association physical association Table 1 and Supplemental figure 1 Also an association with AMFR, only found very recently to interact with VCP (43), was detected (Table I). stub1-mapt 7 19 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 25 unspecified role unspecified role bait bait 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) 43 45 physical association physical association Table 1 and supplemental figure 2 Here we show that CHIP interacts with a domain at the carboxyl terminus of Tau (aa 685 756) that is located downstream of the tubulin-binding domains (aa 559 684). stub1-hspa8 7 25 unspecified role unspecified role bait bait 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) 11 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 53 49 physical association physical association Table 1 and supplemental figure 2 interaction partners Hsp70, Hsc70, and Hsp90 were detected by the GST-CHIP overlay assay (Table I and Supplemental Fig. 2) stub1-hspa1b 7 25 unspecified role unspecified role bait bait 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) 20 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 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 ecoli Escherichia coli (strain K12) 59 56 physical association physical association Table 1 and supplemental figure 2 interaction partners Hsp70, Hsc70, and Hsp90 were detected by the GST-CHIP overlay assay (Table I and Supplemental Fig. 2) stub1-hsp90a 7 25 unspecified role unspecified role bait bait 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) 21 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 ecoli Escherichia coli (strain K12) 66 63 physical association physical association Table 1 and supplemental figure 2 interaction partners Hsp70, Hsc70, and Hsp90 were detected by the GST-CHIP overlay assay (Table I and Supplemental Fig. 2) stub1-ube2q 7 22 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 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 ecoli Escherichia coli (strain K12) 25 unspecified role unspecified role bait bait 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) 72 69 physical association physical association Table 1 and supplemental figure 2 uncharacterized ubiquitin-conjugating enzyme E2Q as well as the disease protein caytaxin (45) interact with CHIP in the overlay screen (Table I). stub1-atcay 7 25 unspecified role unspecified role bait bait 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) 12 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 76 80 physical association physical association Table 1 and supplemental figure 2 uncharacterized ubiquitin-conjugating enzyme E2Q as well as the disease protein caytaxin (45) interact with CHIP in the overlay screen (Table I). bin1-dnm1 7 13 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 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait sh3 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) 86 83 physical association physical association Table 1 and supplemental figure 2 Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). bin1-dlp4 7 15 unspecified role unspecified role bait bait sh3 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) 9 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 90 94 physical association physical association Table 1 and supplemental figure 2 the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners bin1-xrcc4 7 17 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 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait sh3 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) 100 97 physical association physical association Table 1 and supplemental figure 2 the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners bin1-fbp1 7 23 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait sh3 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 Table 1 and supplemental figure 2 the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners vcp-amfr-1 5 18 unspecified role unspecified role bait bait n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 14 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 115 112 physical association physical association Fig 2A, 2B Fig. 2A shows the selective enrichment on the GST-VCP affinity matrix of a 35-kDa carboxyl-terminal His-tagged AMFR fragment from a crude E. coli protein extract. In strong contrast, no AMFR protein was bound to the matrix with the control protein GST. vcp-amfr-2 2 18 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait c terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2C The interaction between VCP and AMFR was also detected by co-immunoprecipitation experiments using transiently transfected COS-1 cells co-expressing full-length VCP and an amino-terminally truncated HA-tagged AMFR fragment. Q9UKV5-1; P55072 transfection vcp-amfr-3 6 14 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 18 unspecified role unspecified role prey prey physical association physical association 2D Co-localization of VCP, labeled with CY3 (red) and AMFR, labeled with Alexa 488 (green), is indicated by the yellow color when the cell is observed under a triple filter. Both proteins co-localized in the cytoplasm as well as in membranous structures in the perinuclear region of COS-1 cells (Fig. 2D) P55072; Q9UKV5-1 transfection vcp-amfr-4 1 14 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 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 ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role bait bait n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Fig 2E, 2F We found that the amino-terminal region of VCP (aa 1 199) is necessary and sufficient for the association with His-AMFR in vitro amfr-vcp 1 18 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 ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role bait bait c terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1783548]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 644-644, sequence length 643 [EBI-1783548]498-644 n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig 2G, 2H The VCP binding region in AMFR was mapped downstream of the Cue domain to the carboxyl terminus of the protein (aa 498�643) (Fig. 2, G and H). stub1-mapt-1 1 19 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 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 ecoli Escherichia coli (strain K12) 25 unspecified role unspecified role bait bait 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 3 As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-hsc70 1 25 unspecified role unspecified role bait bait 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) 11 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 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 3 As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-hsp70 1 25 unspecified role unspecified role bait bait 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) 20 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2010570]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 646-646, sequence length 641 [EBI-2010570]417-646 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 3A As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-hsp90 1 25 unspecified role unspecified role bait bait 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) 21 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 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 3A As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-ube2q-1 1 22 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 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 ecoli Escherichia coli (strain K12) 25 unspecified role unspecified role bait bait 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 3A As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-atcay-1 1 12 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 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 ecoli Escherichia coli (strain K12) 25 unspecified role unspecified role bait bait 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) 176 179 physical association physical association Fig 3A As shown in Fig. 3A the recombinant proteins Tau, Hsc70, Hsp70, Hsp90, E2Q, and caytaxin interact with GST-CHIP in pull-down assays stub1-hsc70-1 8 11 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 25 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 3B stub1-hsp70-1 8 20 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2010524]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 646-646, sequence length 641 [EBI-2010524]417-646 25 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 3B stub1-hsp90a-1 8 21 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 25 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 3B stub1-ube2q-2 8 25 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 22 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 physical association physical association 3B atcay-stub1 8 25 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 12 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 3B chip-ubch5b 4 24 unspecified role unspecified role neutral component neutral component 16 enzyme target enzyme target substrate substrate neutral component neutral component 12 enzyme target enzyme target substrate substrate neutral component neutral component 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 ecoli Escherichia coli (strain K12) 25 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) ubiquitination ubiquitination reaction ubiquitination Fig 3C CHIP efficiently stimulated the ubiquitination of caytaxin, indicating that the protein is a substrate for the E3 ligase activity of CHIP and that the interaction between both proteins could be important for regulation of caytaxin turnover in vivo. The expressedin for E1 and UBcH5b is not known. Ubquitin was also part of the interaction since ubiquitination observed - however source of Ubiquitin unknown. Caytaxin is ubiquitinated. bin1-dlp4-1 8 9 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 15 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 4B Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B) bin1-dlp4-2 1 9 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 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait 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 4A Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B). bin1-xrcc4-1 1 17 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 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait 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 4A Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B). bin1-fbp 1 23 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 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait 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 4A Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B). bin1-fbp-1 8 15 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 23 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 physical association physical association 4B Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B). bin1-xrcc4-2 8 15 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 17 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 physical association physical association 4B Overlay experiments with the SH3 domain of amphiphysin II allowed the identification of the known interaction partner dynamin (Table I). In addition, the proteins Discs large-associated protein 4 (DLP4), XRCC4 (DNA repair protein), and fructose-1,6-bisphosphatase (FBP) (Table I) were identified as novel amphiphysin II interaction partners and verified by in vitro pull-down and two-hybrid assays (Fig. 4, A and B). bin1-dlp4-3 3 15 unspecified role unspecified role prey prey 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 sh3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait 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 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association 4C, 4D We found that in the protein DLP4, besides a proline-rich motif (aa 915�941; Fig. 4E) with a well known PXXP core element (49), also the sequences RRDGYWFLKLLQAET (aa 798�812) and KQRQEARKRLLAAKRAASVRQNSA (aa 948�971) were recognized by the SH3 domain of amphiphysin II (Fig. 4, C and D). bin1-xrcc4-3 3 17 unspecified role unspecified role bait bait 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 chemical synthesis Chemical synthesis (Chemical synthesis) 15 unspecified role unspecified role prey prey 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 sh3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association 4C, 4D Interestingly in XRCC4 arginine- and lysine-rich sequences were found to be critical for the association with the SH3 domain (Fig. 4, C�E). Besides the Arg/Lys-rich sequence in XRCC4 (aa 259�285; Fig. 4E) also the peptide KISRIHLVSEPSITH (aa 4�18) gave a signal above background (Fig. 4, C and D) bin1-fbp-2 3 15 unspecified role unspecified role prey prey 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 sh3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 23 unspecified role unspecified role bait bait region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association 4C, 4D In FBP, the peptides RKARGTGELTQLLNS (aa 22 36) and GTIFGIYRKKSTDEP (aa 130 147) were detected by overlay screens (Fig. 4, C and D).