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 CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. EMBO J. (0261-4189) 2011 imex curation Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. Only protein-protein interactions human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. EMBO J. (0261-4189) 2011 imex curation Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. Only protein-protein interactions rat-primary cerebell Rattus norvegicus primary cerebellar n primary cerebellar n fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. EMBO J. (0261-4189) 2011 imex curation Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. Only protein-protein interactions human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. EMBO J. (0261-4189) 2011 imex curation Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. Only protein-protein interactions rat-brain synaptosom Rattus norvegicus brain_synaptosome brain synaptosome anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. CSN complex controls the stability of selected synaptic proteins via a torsinA-dependent process. EMBO J. (0261-4189) 2011 imex curation Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. Only protein-protein interactions rat-hippocampus_neuron Rattus norvegicus brain-hippocampus_neuron Hippocampal neuron fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Granata A., Koo SJ., Haucke V., Schiavo G., Warner TT. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation nedd8_human NEDD8 Ubiquitin-like protein Nedd8 Neddylin Neural precursor cell expressed developmentally down-regulated protein 8 NEDD8 protein protein human Homo sapiens Human MLIKVKTLTGKEIEIDIEPTDKVERIKERVEEKEGIPPQQQRLIYSGKQMNDEKTAADYKILGGSVLHLVLALRGGGGLRQ DIP protein Q15843 original sequence version: 103 DIP protein Q15843 original sequence version: 149 DIP protein Q15843 original sequence version: 58 DIP protein Q15843 original sequence version: 124 DIP protein Q15843 original sequence version: 95 DIP protein Q15843 original sequence version: 111 DC2FE102BE4725D2 tor1a_human Torsin-1A Torsin family 1 member A Dystonia 1 protein TOR1A DQ2 DYT1 TA Torsin ATPase-1A TORA protein protein human Homo sapiens Human MKLGRAVLGLLLLAPSVVQAVEPISLGLALAGVLTGYIYPRLYCLFAECCGQKRSLSREALQKDLDDNLFGQHLAKKIILNAVFGFINNPKPKKPLTLSLHGWTGTGKNFVSKIIAENIYEGGLNSDYVHLFVATLHFPHASNITLYKDQLQLWIRGNVSACARSIFIFDEMDKMHAGLIDAIKPFLDYYDLVDGVSYQKAMFIFLSNAGAERITDVALDFWRSGKQREDIKLKDIEHALSVSVFNNKNSGFWHSSLIDRNLIDYFVPFLPLEYKHLKMCIRVEMQSRGYEIDEDIVSRVAEEMTFFPKEERVFSDKGCKTVFTKLDYYYDD B69B28D0B4112080 tor1a_rat Torsin-1A Dystonia 1 protein Torsin ATPase 1 Tor1a Dyt1 protein protein rat Rattus norvegicus (Rat) Rat MKLGRATLALLLLVPCVVRAVEPISLGLALAGVLTGYISYPRLYCLFAECCGQKRSLSREALQKDLDNKLFGQHLAKRVILNAVSGFLSNPKPKKPLTLSLHGWTGTGKNFASKIIAENIYEGGLNSDYVHLFVATLHFPHASNITLYKDQLQMWIRGNVSACARSIFIFDEMDKMHAGLIDAIKPFLDYYDVVDEVSYQKAIFIFLSNAGAERITDVALDFWRSGKQREEIKLRDMEHALAVSVFNNKNSGFWHSSLIDRNLIDYFVPFLPLEYKHLKMCIRVEMQSRGYEEDEDIINKVAEEMTFFPKEEKVFSDKGCKTVFTKLDYYLDD mint 8DE5E15F43B67447 ston2_rat Stonin-2 Ston2 protein protein rat Rattus norvegicus (Rat) Rat MTTLDHVIATHQSEWVSFSEEPLFPTPLEGGTEEHFPGLSSSSDRSESSSGENHAVDEGSQDLSHSEQDDSSEKMGLISEAASPPGSPVQPTPDLASAISNWVQFEDDTPWSNTSAPHKETALTLTVPCWTCPSLDSLRRCPLASESSWTTHSEDTSSPSVAPSYTDLQLINAEEQASGRASGTDSTDNSSSLQEDEEVEMETISWWAGSPAMNGHPAVPQVTTARFPSWVTFEDNEVGCPSPTVPSPKKPNAPSAATAGPDVPFNSTGSFKRDRPKSTLMNLSKVQKLDISSLNRPPSVTEAPPWRATNPFLNESLQDIQPSPINPFSAFFEEQERRSQNSSISGTSGKSQRDSLIVIYQDAISFDDSGKSQSHPDAIEKLKQLQIDDPDPVGNAALPDDDPTASVEPDAPSPTSALSQPRDGWPMMLRIPEKKNIMSSRHWGPIYIKLTDSGYLQLYYEQGLEKPFREFKLEICHEVSEPRLQNYDENGRIHSLRIDRVTYKEKKKYQPKPAVAHAAEREQVIKLGTTNYDDFLSFIHAVQDRLMDLPVQSMDLSTVGLNYLEEEITVDIRDEFSGTVSKGDNQILQHHVLTRIHILSFLSGLAECRLGLNDILIKGNEIVSRQDIMPTTTTKWIKLHECRFHGCVDEDVFNSSRVILFNPLDACRFELMRFRTVFAEKTLPFTLRTAASINGAEVEVQSWLRMSPGFSSNRDPLTQVPCENVMVRYPVPSEWVKNFRRESVLGEKSLKAKVNRGASFGSAGASGSEPVMRVTLGTAKYEHAFNSIVWRINRLPDKNSASGHPHCFFCHLELGSDREVPSRFANHVNVEFSMPTTSASKAAVRSISVEDKPDVRKWVNYSAHYSYKVEIEQKKSLKPDFEGEDLENPKECGVQ mint 692E1308E897F35C syt1_rat Synaptotagmin-1 Syt1 Synaptotagmin I p65 protein protein rat Rattus norvegicus (Rat) Rat MVSASHPEALAAPVTTVATLVPHNATEPASPGEGKEDAFSKLKQKFMNELHKIPLPPWALIAIAIVAVLLVVTCCFCVCKKCLFKKKNKKKGKEKGGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK DIP protein P21707 original sequence version: 71 DIP protein P21707 original sequence version: 76 DIP protein P21707 original sequence version: 141 DIP protein P21707 original sequence version: 112 DIP protein P21707 original sequence version: 145 DIP protein P21707 original sequence version: 160 06CE28F04C97A722 snapn_rat SNARE-associated protein Snapin Snapap Synaptosomal-associated protein 25-binding protein Snapin Snap25bp Biogenesis of lysosome-related organelles complex 1 subunit 7 Bloc1s7 protein protein rat Rattus norvegicus (Rat) Rat MAAAGSAAVSGAGTPVAGPTGRDLFAEGLLEFLRPAVQQLDSHVHAVRESQVELREQIDNLATELCRINEDQKVALDLDPYVKKLLNARRRVVLVNNILQNAQERLRRLNHSVAKETARRRAMLDSGVYPPGSPSK C81AC2ABCDCA21FA csn4_rat COP9 signalosome complex subunit 4 Cops4 Csn4 JAB1-containing signalosome subunit 4 protein protein rat Rattus norvegicus (Rat) Rat MAAAVRQDLAQLMNSSGSHKDLAGKYRQILEKAIQLSGTEQLEALKAFVEAMVNENVSLVISRQLLTDFCTHLPNLPDSTAKEVYHFTLEKVQPRVISFEEQVASIRQRLASIYEKEEDWRNAAQVLVGIPLETGQKQYNVDYKLETYLKIARLYLEDDDPVQAEAYINRASLLQNESTNEQLQIHYKVCYARVLDYRRKFIEAAQRYNELSYKTIVHESERLEALKHALHCTILASAGQQRSRMLATLFKDERCQQLAAYGILEKMYLDRIIRGNQLQEFAAMLMPHQKATTADGSSILDRAVIEHNLLSASKLYNNITFEELGALLEIPAAKAEKIASQMITEGRMNGFIDQIDGIVHFETREALPTWDKQIQSLCFQVNNLLEKISQTAPEWTAQAMEAQMAQ mint Component of the COP9 signalosome complex (CSN), a complex involved in various cellular and developmental processes. The CSN complex is an essential regulator of the ubiquitin (Ubl) conjugation pathway by mediating the deneddylation of the cullin subunits of SCF-type E3 ligase complexes, leading to decrease the Ubl ligase activity of SCF. The complex is also involved in phosphorylation of p53/TP53, c-jun/JUN, IkappaBalpha/NFKBIA and IRF8/ICSBP, possibly via its association with CK2 and PKD kinases. CSN-dependent phosphorylation of TP53 and JUN promotes and protects degradation by the Ubl system, respectively B0F34DFC48E8DDA2 ston2_human Stonin-2 Stoned B STON2 STNB STN2 protein protein human Homo sapiens Human MTTLDHVIATHQSEWVSFNEEPPFPAHSQGGTEEHLPGLSSSPDQSESSSGENHVVDGGSQDHSHSEQDDSSEKMGLISEAASPPGSPEQPPPDLASAISNWVQFEDDTPWASTSPPHQETAETALPLTMPCWTCPSFDSLGRCPLTSESSWTTHSEDTSSPSFGCSYTDLQLINAEEQTSGQASGADSTDNSSSLQEDEEVEMEAISWQASSPAMNGHPAPPVTSARFPSWVTFDDNEVSCPLPPVTSPLKPNTPPSASVIPDVPYNSMGSFKKRDRPKSTLMNFSKVQKLDISSLNRTPSVTEASPWRATNPFLNETLQDVQPSPINPFSAFFEEQERRSQNSSISSTTGKSQRDSLIVIYQDAISFDDSSKTQSHSDAVEKLKQLQIDDPDHFGSATLPDDDPVAWIELDAHPPGSARSQPRDGWPMMLRIPEKKNIMSSRHWGPIFVKLTDTGYLQLYYEQGLEKPFREFKLEICHEISEPRLQNYDENGRIHSLRIDRVTYKEKKKYQPKPAVAHTAEREQVIKLGTTNYDDFLSFIHAVQDRLMDLPVLSMDLSTVGLNYLEEEITVDVRDEFSGIVSKGDNQILQHHVLTRIHILSFLSGLAECRLGLNDILVKGNEIVLRQDIMPTTTTKWIKLHECRFHGCVDEDVFHNSRVILFNPLDACRFELMRFRTVFAEKTLPFTLRTATSVNGAEVEVQSWLRMSTGFSANRDPLTQVPCENVMIRYPVPSEWVKNFRRESVLGEKSLKAKVNRGASFGSTSVSGSEPVMRVTLGTAKYEHAFNSIVWRINRLPDKNSASGHPHCFFCHLELGSDREVPSRFANHVNVEFSMPTTSASKASVRSISVEDKTDVRKWVNYSAHYSYQVALGSIWLMLPTPFVHPTTLPLLFLLAMLTMFAW 59F5810DEC540CA3 syt1_human Synaptotagmin-1 Synaptotagmin I p65 SYT1 SYT SVP65 protein protein human Homo sapiens Human MVSESHHEALAAPPVTTVATVLPSNATEPASPGEGKEDAFSKLKEKFMNELHKIPLPPWALIAIAIVAVLLVLTCCFCICKKCLFKKKNKKKGKEKGGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK DIP protein P21579 original sequence version: 146 467F7C58E411AFA9 cul1_human Cullin-1 CUL1 protein protein human Homo sapiens Human MSSTRSQNPHGLKQIGLDQIWDDLRAGIQQVYTRQSMAKSRYMELYTHVYNYCTSVHQSNQARGAGVPPSKSKKGQTPGGAQFVGLELYKRLKEFLKNYLTNLLKDGEDLMDESVLKFYTQQWEDYRFSSKVLNGICAYLNRHWVRRECDEGRKGIYEIYSLALVTWRDCLFRPLNKQVTNAVLKLIEKERNGETINTRLISGVVQSYVELGLNEDDAFAKGPTLTVYKESFESQFLADTERFYTRESTEFLQQNPVTEYMKKAEARLLEEQRRVQVYLHESTQDELARKCEQVLIEKHLEIFHTEFQNLLDADKNEDLGRMYNLVSRIQDGLGELKKLLETHIHNQGLAAIEKCGEAALNDPKMYVQTVLDVHKKYNALVMSAFNNDAGFVAALDKACGRFINNNAVTKMAQSSSKSPELLARYCDSLLKKSSKNPEEAELEDTLNQVMVVFKYIEDKDVFQKFYAKMLAKRLVHQNSASDDAEASMISKLKQACGFEYTSKLQRMFQDIGVSKDLNEQFKKHLTNSEPLDLDFSIQVLSSGSWPFQQSCTFALPSELERSYQRFTAFYASRHSGRKLTWLYQLSKGELVTNCFKNRYTLQASTFQMAILLQYNTEDAYTVQQLTDSTQIKMDILAQVLQILLKSKLLVLEDENANVDEVELKPDTLIKLYLGYKNKKLRVNINVPMKTEQKQEQETTHKNIEEDRKLLIQAAIVRIMKMRKVLKHQQLLGEVLTQLSSRFKPRVPVIKKCIDILIEKEYLERVDGEKDTYSYLA DIP protein Q13616 original sequence version: 144 DIP protein Q13616 original sequence version: 133 DIP protein Q13616 original sequence version: null DIP protein Q13616 original sequence version: 135 DIP protein Q13616 original sequence version: 98 DIP protein Q13616 original sequence version: 110 DIP protein Q13616 original sequence version: 146 DIP protein Q13616 original sequence version: 148 DIP protein Q13616 original sequence version: 100 DIP protein Q13616 original sequence version: 85 DIP protein Q13616 original sequence version: 114 DIP protein Q13616 original sequence version: 119 DIP protein Q13616 original sequence version: 136 DIP protein Q13616 original sequence version: 117 DIP protein Q13616 original sequence version: 99 DIP protein Q13616 original sequence version: 108 6625A1FFA7799BBA csn4_human COP9 signalosome complex subunit 4 COPS4 CSN4 JAB1-containing signalosome subunit 4 protein protein human Homo sapiens Human MAAAVRQDLAQLMNSSGSHKDLAGKYRQILEKAIQLSGAEQLEALKAFVEAMVNENVSLVISRQLLTDFCTHLPNLPDSTAKEIYHFTLEKIQPRVISFEEQVASIRQHLASIYEKEEDWRNAAQVLVGIPLETGQKQYNVDYKLETYLKIARLYLEDDDPVQAEAYINRASLLQNESTNEQLQIHYKVCYARVLDYRRKFIEAAQRYNELSYKTIVHESERLEALKHALHCTILASAGQQRSRMLATLFKDERCQQLAAYGILEKMYLDRIIRGNQLQEFAAMLMPHQKATTADGSSILDRAVIEHNLLSASKLYNNITFEELGALLEIPAAKAEKIASQMITEGRMNGFIDQIDGIVHFETREALPTWDKQIQSLCFQVNNLLEKISQTAPEWTAQAMEAQMAQ DIP protein Q9BT78 original sequence version: 72 DIP protein Q9BT78-1 original sequence version: 115 DIP protein Q9BT78 original sequence version: 132 DIP protein Q9BT78 original sequence version: 110 4EFF9079058EB609 cops4-tor1a 1 Csn4 16 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell TA 7 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f2C homomint mint immunostaining with CSN4 and TA antibodies showed a partial co-localisation of both proteins on discrete puncta along neurites in SH-SY5Y cells (Figure 2C) and in primary cerebellar neurons (Figure 2D) cops4-syt1 1 Csn4 16 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Syt1 14 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization homomint mint sf1f Notably, CSN4 was seen to partially overlap with SV markers, such as Syt1 in SH-SY5Y cells (Supplementary Figure 1A F) and in primary hippocampal neurons (Supplementary Figure 1G L). cops4-dyt1 4 TA 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Csn4 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Snapin 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f2b homomint mint ston2-cops4 4 TA 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Csn4 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stonin2 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association f5b mint cops4-dyt1-1 2 TA 8 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Csn4 12 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f2d mint immunostaining with CSN4 and TA antibodies showed a partial co-localisation of both proteins on discrete puncta along neurites in SH-SY5Y cells (Figure 2C) and in primary cerebellar neurons (Figure 2D) cops4-syt1-1 5 Csn4 12 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Syt1 10 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization homomint mint Notably, CSN4 was seen to partially overlap with SV markers, such as Syt1 in SH-SY5Y cells (Supplementary Figure 1A F) and in primary hippocampal neurons (Supplementary Figure 1G L). sf1l nedd8-cul1-1 3 NEDD8 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level CUL1 15 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level stonin2 13 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association homomint mint sf5 The intensity of the neddylated stonin 2 bands is stronger in cells overexpressing E-TA (E) compared with control cells (Ctl) and cells expressing wt- TA (wt), whilst the level of cullin 1 (CUL-1), a known neddylated target of CSN complex, remained unchanged. ston2-nedd8-1 3 stonin2 13 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level NEDD8 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f5d homomint mint Blotting with specific antibodies against NEDD8 and ubiquitin showed that the upper bands recovered in anti-stonin 2 immunoprecipitation samples correspond to neddylated and polyubiquitinated form of stonin 2 (Figure 5D).