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/ Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Cell (0092-8674) 2014 imex curation Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro neddylase assay neddylase assay tag fluorescence tag visualisation by fluorescence tag fluorescence Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. Details of constructs (tags, length ad expression system) confused as they refer back to multiple papers which contain different version of the proteins. If in doubt, I've matched to the crystal, unless its clear they have used something different. Interactions involving the cyclin E peptide not included as I cannot work out what this is. Gave up on Figs 5-7 - too complicated to capture. Pulse-chase assays were used to monitoring the ligation reaction Indicated E2-Ubiquitin-like protein (UBL) complexes were formed in the pulse reaction. After quenching the pulse, UBL was “chased” from E2 by adding additional proteins or protein complexes. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org Accepted 2014-NOV-24 by MGT Cell (0092-8674) 2014 Only protein-protein interactions imex curation Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Cell (0092-8674) 2014 imex curation Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. It is not clear if the GST tags on Cullin1 and Nedd8 have been retained, or not. Length and expression vector should be OK, as taken from PDBe - tags still open to question, especially as sometimes cleaved off in crystals in other papers. So, more of a best guess than necessarily fact. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org Accepted 2014-NOV-24 by MGT Cell (0092-8674) 2014 Only protein-protein interactions imex curation Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Structure of a RING E3 trapped in action reveals ligation mechanism for the ubiquitin-like protein NEDD8. Cell (0092-8674) 2014 imex curation Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro ubiquitinase assay ubiquitinase assay ubiquitinase assay tag fluorescence tag visualisation by fluorescence tag fluorescence Scott D.C., Sviderskiy V.O., Monda J.K., Lydeard J.R., Cho S.E., Harper J.W., Schulman B.A. Details of constructs (tags, length ad expression system) confused as they refer back to multiple papers which contain different version of the proteins. Where I can I've matched to the crystal, unless its clear they have used something different. Pulse-chase assays were used to monitoring the ligation reaction Indicated E2-Ubiquitin-like protein (UBL) complexes were formed in the pulse reaction. After quenching the pulse, UBL was “chased” from E2 by adding additional proteins or protein complexes. Details of some constructs (e.g. tags, length and expression system) are not clear; when in doubt these have been matched to constructs used in the crystal. brenda.schulman@stjude.org Accepted 2014-NOV-24 by MGT Cell (0092-8674) 2014 Only protein-protein interactions imex curation rbx1_human E3 ubiquitin-protein ligase RBX1 RBX1 RNF75 ROC1 Regulator of cullins 1 RING finger protein 75 Protein ZYP RING-box protein 1 E3 ubiquitin-protein transferase RBX1 protein protein human Homo sapiens Human MAAAMDVDTPSGTNSGAGKKRFEVKKWNAVALWAWDIVVDNCAICRNHIMDLCIECQANQASATSEECTVAWGVCNHAFHFHCISRWLKTRQVCPLDNREWEFQKYGH DIP protein P62877 original sequence version: 120 DIP protein P62877 original sequence version: 115 DIP protein P62877 original sequence version: 78 DIP protein P62877 original sequence version: 113 DIP protein P62877 original sequence version: 75 DIP protein P62877 original sequence version: 86 DIP protein P62877 original sequence version: 136 DIP protein P62877 original sequence version: 126 DIP protein P62877 original sequence version: 90 DIP protein P62877 original sequence version: 114 30FC5ADF66096C0E 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 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 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 ubiq_human UBIQ_HUMAN protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG dcnl1_human DCN1-like protein 1 DCUN1D1 DCUN1L1 RP42 SCCRO Defective in cullin neddylation protein 1-like protein 1 DCUN1 domain-containing protein 1 Squamous cell carcinoma-related oncogene DCN1 protein protein human Homo sapiens Human MNKLKSSQKDKVRQFMIFTQSSEKTAVSCLSQNDWKLDVATDNFFQNPELYIRESVKGSLDRKKLEQLYNRYKDPQDENKIGIDGIQQFCDDLALDPASISVLIIAWKFRAATQCEFSKQEFMDGMTELGCDSIEKLKAQIPKMEQELKEPGRFKDFYQFTFNFAKNPGQKGLDLEMAIAYWNLVLNGRFKFLDLWNKFLLEHHKRSIPKDTWNLLLDFSTMIADDMSNYDEEGAWPVLIDDFVEFARPQIAGTKSTTV DIP protein Q96GG9 original sequence version: 94 F3709235E0610C54 ubc12_human NEDD8-conjugating enzyme Ubc12 UBE2M UBC12 Ubiquitin-conjugating enzyme E2 M NEDD8 carrier protein protein protein human Homo sapiens Human MIKLFSLKQQKKEEESAGGTKGSSKKASAAQLRIQKDINELNLPKTCDISFSDPDDLLNFKLVICPDEGFYKSGKFVFSFKVGQGYPHDPPKVKCETMVYHPNIDLEGNVCLNILREDWKPVLTINSIIYGLQYLFLEPNPEDPLNKEAAEVLQNNRRLFEQNVQRSMRGGYIGSTYFERCLK DIP protein P61081 original sequence version: 108 DIP protein P61081 original sequence version: 84 DIP protein P61081 original sequence version: 106 E3C288CA6A98BC5C cul1-nedd8-1 1 4 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P62877:p.Arg46Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A Wed May 02 20:24:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg46ala' Tue Jun 30 15:44:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg46Ala' P62877:p.Asn98Arg mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain N R Wed May 02 17:08:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98arg' Tue Jun 30 16:51:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Arg' P62877:p.Ile37Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain I A Wed May 02 18:43:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile37ala' Tue Jun 30 20:25:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile37Ala' P62877:p.Ile37_Val38delinsGlyGly mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain IV GG Thu Jul 07 12:16:42 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ile37_val38gly Fri Jul 22 11:11:22 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ile37gly,val38gly' Wed May 02 16:14:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile_val37-38gly_gly' Tue Jun 30 16:44:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile37_Val38delinsGlyGly' P62877:p.Trp35Asp mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain W D Wed May 02 19:00:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp35asp' Tue Jun 30 19:43:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp35Asp' ecoli Escherichia coli (strain K12) 10 ancillary ancillary neutral component neutral component P61081:p.Asn113Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain N A Wed May 02 18:57:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn113ala' Tue Jun 30 18:16:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn113Ala' 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 P61081:p.Arg116Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A Wed May 02 14:25:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg116ala' Tue Jun 30 17:01:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg116Ala' ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q13616:p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain VV EK certain certain sequence position certain certain certain sequence position certain Y K Thu Jul 07 12:07:23 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu421glu_val451glu_val452lys_tyr455lys Fri Jul 22 10:51:24 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val451glu,val452lys,tyr455lys' Wed May 02 18:11:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val_val451-452glu_lys,tyr455lys' Tue Jun 30 18:56:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys]' Q13616:p.Lys720Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:06:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys720ala' Tue Jun 30 19:47:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys720Ala' ecoli Escherichia coli (strain K12) 7 ancillary ancillary neutral component neutral component 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 region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q15843:p.Ile44Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain I A Wed May 02 15:26:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile44ala' Tue Jun 30 20:13:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile44Ala' Q15843:p.Leu8Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain L A Wed May 02 18:21:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu8ala' Tue Jun 30 19:39:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu8Ala' Q15843:p.Leu8_Thr9delinsAlaAla mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain LT AA Thu Jul 07 12:17:15 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu8_thr9ala Fri Jul 22 11:07:25 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu8ala,thr9ala' Wed May 02 19:09:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_thr8-9ala_ala' Tue Jun 30 16:52:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu8_Thr9delinsAlaAla' Q15843:p.Glu32Asp mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain E D Wed May 02 15:27:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu32asp' Tue Jun 30 17:31:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu32Asp' Q15843:p.Glu31_Glu32delinsGlnAsp mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain EE QD Thu Jul 07 12:16:12 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was glu31gln_glu32asp Fri Jul 22 11:10:16 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu31gln,glu32asp' Wed May 02 15:15:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu_glu31-32gln_asp' Tue Jun 30 17:06:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu31_Glu32delinsGlnAsp' ecoli Escherichia coli (strain K12) neddylation neddylation reaction neddylation Fig. 1B, 1D, 1G, 3D, 3H, 3K, 3L, 4D, 4E, 4G. Supp. Fig. 1C, 1D, Subsequent analysis of the product by western blot confirmed the fluorescent tag on NEDD8 had no affect on reaction rate. Introduction of lysines surrounding the CUL1 lys720ala substitution, or appending Gly-Gly-Lys to the C terminus fails to rescue CUL1 neddylation. UBC12 tyr130leu mutations rescues NEDD8 ile44ala mutation. cul1-rbx1-9 3 8 ancillary ancillary neutral component neutral component region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ile44ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain I A Thu Jul 07 12:17:35 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ile44ala asp32glu mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain D E Thu Jul 07 12:16:14 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was asp32glu gln31glu,asp32glu mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain QD EE Thu Jul 07 12:16:58 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was gln31_asp32glu 5 ancillary ancillary neutral component neutral component P62837:p.Ser108Leu mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain S L Wed May 02 16:27:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser108leu' Tue Jun 30 20:08:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser108Leu' ecoli Escherichia coli (strain K12) 4 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P62877:p.Asn98Arg mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain N R Wed May 02 19:10:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98arg' Tue Jun 30 18:53:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Arg' P62877:p.Asn98Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain N A Wed May 02 20:08:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98ala' Tue Jun 30 20:23:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Ala' P62877:p.Trp35Asp mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain W D Wed May 02 15:03:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp35asp' Tue Jun 30 17:14:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp35Asp' ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component Q13616:p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain VV EK certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain Y K Thu Jul 07 12:17:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu421glu_val451glu_val452_lys_tyr455lys Fri Jul 22 10:58:52 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val451glu,val452lys,tyr455lys' Wed May 02 19:21:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val_val451-452glu_lys,tyr455lys' Tue Jun 30 19:12:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys]' c-terminal 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) ubiquitination ubiquitination reaction ubiquitination Fig. 1C, 3E, 4F, 4H RBX1 promotingted ubiquitin ligation from UBCH5 to CUL1 more slowly that is did ligation from UBC12-NEDD8. UBCH5B ser108leu mutations rescue UBIQ ile44ala mutations cul1-rbx1-10 3 4 enzyme enzyme neutral component neutral component P62877:p.Asn98Arg mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain N R Wed May 02 16:33:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98arg' Tue Jun 30 15:43:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Arg' 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) 8 ancillary ancillary neutral component neutral component arg72ala mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain R A Thu Jul 07 12:15:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was arg72ala 10 ancillary ancillary neutral component neutral component 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) 6 enzyme target enzyme target substrate substrate neutral component neutral component Q13616:p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain VV EK certain certain sequence position certain certain certain sequence position certain Y K Thu Jul 07 12:15:39 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu421glu_val451glu_val452_lys_tyr455lys Fri Jul 22 11:10:26 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val451glu,val452lys,tyr455lys' Wed May 02 16:24:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val_val451-452glu_lys,tyr455lys' Tue Jun 30 18:10:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys]' c-terminal 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) ubiquitination ubiquitination reaction ubiquitination Fig. 1D cul1-rbx1-11 1 4 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P62877:p.Asn98Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain N A Wed May 02 18:59:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98ala' Tue Jun 30 18:15:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Ala' P62877:p.Asn98Arg mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain N R Wed May 02 19:16:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn98arg' Tue Jun 30 20:13:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn98Arg' ecoli Escherichia coli (strain K12) 7 ancillary ancillary neutral component neutral component region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q15843:p.Ala72Arg mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain A R Wed May 02 16:19:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala72arg' Tue Jun 30 18:30:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala72Arg' 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) 5 ancillary ancillary neutral component neutral component ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component Q13616:p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain VV EK certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain Y K Thu Jul 07 12:16:44 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu421glu_val451glu_val452_lys_tyr455lys Fri Jul 22 10:58:27 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val451glu,val452lys,tyr455lys' Wed May 02 15:30:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val_val451-452glu_lys,tyr455lys' Tue Jun 30 17:13:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys]' c-terminal 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) neddylation neddylation reaction neddylation Fig. 1E RBX1 promotingted ubiquitin ligation from UBCH5 to CUL1 more slowly that is did ligation from UBC12-NEDD8. cul1-dcun1d1 2 9 unspecified role unspecified role neutral component neutral component 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) 4 unspecified role unspecified role neutral component neutral component 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) 10 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P61081:p.[Asn103Ser;Cys111Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain N S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 14:52:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn103ser,cys111ser' Tue Jun 30 17:49:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn103Ser;Cys111Ser]' c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 7 unspecified role unspecified role neutral component neutral component 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) 6 unspecified role unspecified role neutral component neutral component Q13616:p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys;Lys720Arg] mutation mutation certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain Y K certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain VV EK Thu Jul 07 12:15:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu421glu_val451glu_val452_lys_tyr455lys_lys720arg Fri Jul 22 11:09:58 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val451glu,val452lys,tyr455lys,lys720arg' Wed May 02 19:45:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu421glu,val_val451-452glu_lys,tyr455lys,lys720arg' Tue Jun 30 20:20:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu421Glu;Val451_Val452delinsGluLys;Tyr455Lys;Lys720Arg]' 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 3.1A Rworking = 0.23 Rfree = 0.28 Supp. Table 1B