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/ http://www.ebi.ac.uk/ kozlov-2004-2 Structural basis of ligand recognition by PABC, a highly specific peptide-binding domain found in poly(A)-binding protein and a HECT ubiquitin ligase. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Kozlov G., De Crescenzo G., Lim N.S., Siddiqui N., Fantus D., Kahvejian A., Trempe J.F., Elias D., Ekiel I., Sonenberg N., O'Connor-McCourt M., Gehring K. kalle.gehring@mcgill.ca EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation kozlov-2004-3 Structural basis of ligand recognition by PABC, a highly specific peptide-binding domain found in poly(A)-binding protein and a HECT ubiquitin ligase. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Kozlov G., De Crescenzo G., Lim N.S., Siddiqui N., Fantus D., Kahvejian A., Trempe J.F., Elias D., Ekiel I., Sonenberg N., O'Connor-McCourt M., Gehring K. kalle.gehring@mcgill.ca EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation kozlov-2004-4 Structural basis of ligand recognition by PABC, a highly specific peptide-binding domain found in poly(A)-binding protein and a HECT ubiquitin ligase. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Kozlov G., De Crescenzo G., Lim N.S., Siddiqui N., Fantus D., Kahvejian A., Trempe J.F., Elias D., Ekiel I., Sonenberg N., O'Connor-McCourt M., Gehring K. kalle.gehring@mcgill.ca EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation kozlov-2004-1 Structural basis of ligand recognition by PABC, a highly specific peptide-binding domain found in poly(A)-binding protein and a HECT ubiquitin ligase. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Kozlov G., De Crescenzo G., Lim N.S., Siddiqui N., Fantus D., Kahvejian A., Trempe J.F., Elias D., Ekiel I., Sonenberg N., O'Connor-McCourt M., Gehring K. kalle.gehring@mcgill.ca EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation paip2_human Polyadenylate-binding protein-interacting protein 2 PAIP2 PAIP2A HSPC218 protein protein human Homo sapiens Human MKDPSRSSTSPSIINEDVIINGHSHEDDNPFAEYMWMENEEEFNRQIEEELWEEEFIERCFQEMLEEEEEHEWFIPARDLPQTMDQIQDQFNDLVISDGSSLEDLVVKSNLNPNAKEFVPGVKYGNI pabp1_human Polyadenylate-binding protein 1 PABPC1 PAB1 PABP1 PABPC2 PABP protein protein human Homo sapiens Human MNPSAPSYPMASLYVGDLHPDVTEAMLYEKFSPAGPILSIRVCRDMITRRSLGYAYVNFQQPADAERALDTMNFDVIKGKPVRIMWSQRDPSLRKSGVGNIFIKNLDKSIDNKALYDTFSAFGNILSCKVVCDENGSKGYGFVHFETQEAAERAIEKMNGMLLNDRKVFVGRFKSRKEREAELGARAKEFTNVYIKNFGEDMDDERLKDLFGKFGPALSVKVMTDESGKSKGFGFVSFERHEDAQKAVDEMNGKELNGKQIYVGRAQKKVERQTELKRKFEQMKQDRITRYQGVNLYVKNLDDGIDDERLRKEFSPFGTITSAKVMMEGGRSKGFGFVCFSSPEEATKAVTEMNGRIVATKPLYVALAQRKEERQAHLTNQYMQRMASVRAVPNPVINPYQPAPPSGYFMAAIPQTQNRAAYYPPSQIAQLRPSPRWTAQGARPHPFQNMPGAIRPAAPRPPFSTMRPASSQVPRVMSTQRVANTSTQTMGPRPAAAAAAATPAVRTVPQYKYAAGVRNPQQHLNAQPQVTMQQPAVHVQGQEPLTASMLASAPPQEQKQMLGERLFPLIQAMHPTLAGKITGMLLEIDNSELLHMLESPESLRSKVDEAVAVLQAHQAKEAAQKAVNSATGVPTV paip1_human Polyadenylate-binding protein-interacting protein 1 PAIP1 protein protein human Homo sapiens Human MSDGFDRAPGAGRGRSRGLGRGGGGPEGGGFPNGAGPAERARHQPPQPKAPGFLQPPPLRQPRTTPPPGAQCEVPASPQRPSRPGALPEQTRPLRAPPSSQDKIPQQNSESAMAKPQVVVAPVLMSKLSVNAPEFYPSGYSSSYTESYEDGCEDYPTLSEYVQDFLNHLTEQPGSFETEIEQFAETLNGCVTTDDALQELVELIYQQATSIPNFSYMGARLCNYLSHHLTISPQSGNFRQLLLQRCRTEYEVKDQAAKGDEVTRKRFHAFVLFLGELYLNLEIKGTNGQVTRADILQVGLRELLNALFSNPMDDNLICAVKLLKLTGSVLEDAWKEKGKMDMEEIIQRIENVVLDANCSRDVKQMLLKLVELRSSNWGRVHATSTYREATPENDPNYFMNEPTFYTSDGVPFTAADPDYQEKYQELLEREDFFPDYEENGTDLSGAGDPYLDDIDDEMDPEIEEAYEKFCLESERKRKQ erf3b_human Eukaryotic peptide chain release factor GTP-binding subunit ERF3B GSPT2 ERF3B G1 to S phase transition protein 2 homolog protein protein human Homo sapiens Human MDSGSSSSDSAPDCWDQVDMESPGSAPSGDGVSSAVAEAQREPLSSAFSRKLNVNAKPFVPNVHAAEFVPSFLRGPTQPPTLPAGSGSNDETCTGAGYPQGKRMGRGAPVEPSREEPLVSLEGSNSAVTMELSEPVVENGEVEMALEESWEHSKEVSEAEPGGGSSGDSGPPEESGQEMMEEKEEIRKSKSVIVPSGAPKKEHVNVVFIGHVDAGKSTIGGQIMFLTGMVDKRTLEKYEREAKEKNRETWYLSWALDTNQEERDKGKTVEVGRAYFETERKHFTILDAPGHKSFVPNMIGGASQADLAVLVISARKGEFETGFEKGGQTREHAMLAKTAGVKHLIVLINKMDDPTVNWSIERYEECKEKLVPFLKKVGFSPKKDIHFMPCSGLTGANIKEQSDFCPWYTGLPFIPYLDNLPNFNRSIDGPIRLPIVDKYKDMGTVVLGKLESGSIFKGQQLVMMPNKHNVEVLGILSDDTETDFVAPGENLKIRLKGIEEEEILPGFILCDPSNLCHSGRTFDVQIVIIEHKSIICPGYNAVLHIHTCIEEVEITALISLVDKKSGEKSKTRPRFVKQDQVCIARLRTAGTICLETFKDFPQMGRFTLRDEGKTIAIGKVLKLVPEKD pabpc1-paip2-1 1448971 n/a 1448975 unspecified role unspecified role neutral component neutral component lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 unspecified role unspecified role neutral component neutral component pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1448983 1448981 direct interaction direct interaction false false false 1448971 Fig. 5 Temp = 25C Kd values (molar) were measured at different temperatures: 10C: 7.9 x 10E-8; 15C: 1.4 x 10E-7; 20C: 2.1 x 10E-7; 25C: 3.5 x 10E-7; 30C: 5.6 x 10E-7. pabpc1-paip1-5 1448971 n/a 1448976 unspecified role unspecified role neutral component neutral component pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448977 unspecified role unspecified role neutral component neutral component lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1448988 1448986 direct interaction direct interaction false false false 1448971 Fig. 5b, Fig. 5c Temp = 25C Kd values (molar) were measured at different temperatures: 10C: 8.2 x 10E-7; 15C: 9.4 x 10E-7; 20C: 1.3 x 10E-6; 25C: 1.5 x 10E-6. gspt2-pabpc1 1448971 n/a 1448978 unspecified role unspecified role neutral component neutral component lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 unspecified role unspecified role neutral component neutral component pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1448993 1448991 direct interaction direct interaction false false false 1448971 Fig. 5b, Fig. 5c Temp = 25C Kd values (molar) were measured at different temperatures: 10C: 3.9 x 10E-7; 15C: 7.3 x 10E-7; 20C: 9.2 x 10E-7; 25C: 1.6 x 10E-6; 30C: 2.4 x 10E-6. pabpc1-paip1-6 1448972 n/a 1448976 unspecified role unspecified role bait bait pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448977 unspecified role unspecified role prey prey lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1448998 1448996 direct interaction direct interaction false false false 1448972 Fig. 6 Temp = 25C pabpc1-gspt2 1448972 n/a 1448978 unspecified role unspecified role prey prey lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 unspecified role unspecified role bait bait pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1449003 1449001 direct interaction direct interaction false false false 1448972 Fig. 6c Temp = 25C Kd (molar) for shorter eRF3 peptides: residues 47-65: 3.9 x 10E-6; residues 56-74: 3.1 x 10E-6. pabpc1-paip2-2 1448972 n/a 1448976 unspecified role unspecified role bait bait pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448975 unspecified role unspecified role prey prey Q9BPZ3:p.Leu111Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Pro113Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Ala115Leu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Lys116Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Glu117Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Phe118Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q9BPZ3:p.Pro120Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) 1449015 1449006 direct interaction direct interaction false false false 1448972 Fig. 6c Temp = 25C Kd values (molar) were measured for peptides of different length: residues 106-127: 1.6 x 10E-7; residues 105-120: 3.0 x 10E-6; residues 105-118: 2.6 x 10E-4; residues 109-125: 1.9 x 10E-7. Kd values (molar) were measured for different mutants of the 109-125 peptide: leu111ala: 2.3 x 10E-4; pro113ala: 7.4 x 10E-7; ala115leu: 1.4 x 10E-5; lys116ala: 4.8 x 10E-7; glu117ala: 3.5 x 10E-7; phe118ala: >10E-3; pro120ala: 3.9 x 10E-6. pabpc1-paip1-7 1448973 n/a 1448977 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 false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 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 false pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false P11940:p.Phe567Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P11940:p.Lys580Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false Fig. 4a pabpc1-paip2-3 1448973 n/a 1448975 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 unspecified role unspecified role bait bait pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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 false P11940:p.Phe567Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P11940:p.Lys580Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false Fig. 4b pabpc1-gspt2-1 1448973 n/a 1448978 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 false ecobd Escherichia coli (strain B / BL21-DE3) n/a 1448976 unspecified role unspecified role bait bait pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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 false P11940:p.Phe567Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false Fig. 4c pabpc1-paip2 1448974 n/a 1448975 unspecified role unspecified role neutral component neutral component lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 n/a 1448976 unspecified role unspecified role neutral component neutral component pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 1449044 1449041 direct interaction direct interaction false false false Fig. 1b, Fig. 1c, Fig. 2a, Fig. 2c, Fig. 3b, Fig. 3c pabpc1-paip1-4 1448974 n/a 1448976 unspecified role unspecified role neutral component neutral component pabp_hyd domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 n/a 1448977 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false lig_pam2_1 motif sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 1449050 1449047 direct interaction direct interaction false false false Fig 1d, Fig. 2b, Fig. 2d, Fig. 3d