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/ pfaff-2013-1 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro peptide array peptide array western blot western blot Immuno blot Immuno blot Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-2 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-3 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro pull down pull down weight by staining molecular weight estimation by staining weight by staining Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-4 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro pull down pull down ms participant Identification by mass spectrometry ms participant Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-5 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-6 Structural features of Argonaute-GW182 protein interactions. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-7 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. Cross linker was:- disuccinimidyl suberate niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. pfaff-2013-8 Structural features of Argonaute-GW182 protein interactions. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Human Ago2 appears to have been expressed in both Sf21 and High five insect cells, although which form of expression was used, and when, is not stated. Saturation transfer difference (STD) NMR niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. ago2_human Protein argonaute-2 AGO2 Argonaute RISC catalytic component 2 EIF2C2 Eukaryotic translation initiation factor 2C 2 PAZ Piwi domain protein Protein slicer protein protein human Homo sapiens Human MYSGAGPALAPPAPPPPIQGYAFKPPPRPDFGTSGRTIKLQANFFEMDIPKIDIYHYELDIKPEKCPRRVNREIVEHMVQHFKTQIFGDRKPVFDGRKNLYTAMPLPIGRDKVELEVTLPGEGKDRIFKVSIKWVSCVSLQALHDALSGRLPSVPFETIQALDVVMRHLPSMRYTPVGRSFFTASEGCSNPLGGGREVWFGFHQSVRPSLWKMMLNIDVSATAFYKAQPVIEFVCEVLDFKSIEEQQKPLTDSQRVKFTKEIKGLKVEITHCGQMKRKYRVCNVTRRPASHQTFPLQQESGQTVECTVAQYFKDRHKLVLRYPHLPCLQVGQEQKHTYLPLEVCNIVAGQRCIKKLTDNQTSTMIRATARSAPDRQEEISKLMRSASFNTDPYVREFGIMVKDEMTDVTGRVLQPPSILYGGRNKAIATPVQGVWDMRNKQFHTGIEIKVWAIACFAPQRQCTEVHLKSFTEQLRKISRDAGMPIQGQPCFCKYAQGADSVEPMFRHLKNTYAGLQLVVVILPGKTPVYAEVKRVGDTVLGMATQCVQMKNVQRTTPQTLSNLCLKINVKLGGVNNILLPQGRPPVFQQPVIFLGADVTHPPAGDGKKPSIAAVVGSMDAHPNRYCATVRVQQHRQEIIQDLAAMVRELLIQFYKSTRFKPTRIIFYRDGVSEGQFQQVLHHELLAIREACIKLEKDYQPGITFIVVQKRHHTRLFCTDKNERVGKSGNIPAGTTVDTKITHPTEFDFYLCSHAGIQGTSRPSHYHVLWDDNRFSSDELQILTYQLCHTYVRCTRSVSIPAPAYYAHLVAFRARYHLVDKEHDSAEGSHTSGQSNGRDHQALAKAVQVHQDTLRTMYFA tnr6b_human Trinucleotide repeat-containing gene 6B protein TNRC6B KIAA1093 protein protein human Homo sapiens Human MREKEQEREEQLMEDKKRKKEDKKKKEATQKVTEQKTKVPEVTKPSLSQPTAASPIGSSPSPPVNGGNNAKRVAVPNGQPPSAARYMPREVPPRFRCQQDHKVLLKRGQPPPPSCMLLGGGAGPPPCTAPGANPNNAQVTGALLQSESGTAPDSTLGGAAASNYANSTWGSGASSNNGTSPNPIHIWDKVIVDGSDMEEWPCIASKDTESSSENTTDNNSASNPGSEKSTLPGSTTSNKGKGSQCQSASSGNECNLGVWKSDPKAKSVQSSNSTTENNNGLGNWRNVSGQDRIGPGSGFSNFNPNSNPSAWPALVQEGTSRKGALETDNSNSSAQVSTVGQTSREQQSKMENAGVNFVVSGREQAQIHNTDGPKNGNTNSLNLSSPNPMENKGMPFGMGLGNTSRSTDAPSQSTGDRKTGSVGSWGAARGPSGTDTVSGQSNSGNNGNNGKEREDSWKGASVQKSTGSKNDSWDNNNRSTGGSWNFGPQDSNDNKWGEGNKMTSGVSQGEWKQPTGSDELKIGEWSGPNQPNSSTGAWDNQKGHPLPENQGNAQAPCWGRSSSSTGSEVGGQSTGSNHKAGSSDSHNSGRRSYRPTHPDCQAVLQTLLSRTDLDPRVLSNTGWGQTQIKQDTVWDIEEVPRPEGKSDKGTEGWESAATQTKNSGGWGDAPSQSNQMKSGWGELSASTEWKDPKNTGGWNDYKNNNSSNWGGGRPDEKTPSSWNENPSKDQGWGGGRQPNQGWSSGKNGWGEEVDQTKNSNWESSASKPVSGWGEGGQNEIGTWGNGGNASLASKGGWEDCKRSPAWNETGRQPNSWNKQHQQQQPPQQPPPPQPEASGSWGGPPPPPPGNVRPSNSSWSSGPQPATPKDEEPSGWEEPSPQSISRKMDIDDGTSAWGDPNSYNYKNVNLWDKNSQGGPAPREPNLPTPMTSKSASVWSKSTPPAPDNGTSAWGEPNESSPGWGEMDDTGASTTGWGNTPANAPNAMKPNSKSMQDGWGESDGPVTGARHPSWEEEEDGGVWNTTGSQGSASSHNSASWGQGGKKQMKCSLKGGNNDSWMNPLAKQFSNMGLLSQTEDNPSSKMDLSVGSLSDKKFDVDKRAMNLGDFNDIMRKDRSGFRPPNSKDMGTTDSGPYFEKLTLPFSNQDGCLGDEAPCSPFSPSPSYKLSPSGSTLPNVSLGAIGTGLNPQNFAARQGGSHGLFGNSTAQSRGLHTPVQPLNSSPSLRAQVPPQFISPQVSASMLKQFPNSGLSPGLFNVGPQLSPQQIAMLSQLPQIPQFQLACQLLLQQQQQQQLLQNQRKISQAVRQQQEQQLARMVSALQQQQQQQQRQPGMKHSPSHPVGPKPHLDNMVPNALNVGLPDLQTKGPIPGYGSGFSSGGMDYGMVGGKEAGTESRFKQWTSMMEGLPSVATQEANMHKNGAIVAPGKTRGGSPYNQFDIIPGDTLGGHTGPAGDSWLPAKSPPTNKIGSKSSNASWPPEFQPGVPWKGIQNIDPESDPYVTPGSVLGGTATSPIVDTDHQLLRDNTTGSNSSLNTSLPSPGAWPYSASDNSFTNVHSTSAKFPDYKSTWSPDPIGHNPTHLSNKMWKNHISSRNTTPLPRPPPGLTNPKPSSPWSSTAPRSVRGWGTQDSRLASASTWSDGGSVRPSYWLVLHNLTPQIDGSTLRTICMQHGPLLTFHLNLTQGTALIRYSTKQEAAKAQTALHMCVLGNTTILAEFATDDEVSRFLAQAQPPTPAATPSAPAAGWQSLETGQNQSDPVGPALNLFGGSTGLGQWSSSAGGSSGADLAGASLWGPPNYSSSLWGVPTVEDPHRMGSPAPLLPGDLLGGGSDSI ago1_human Protein argonaute-1 AGO1 Argonaute RISC catalytic component 1 EIF2C1 Eukaryotic translation initiation factor 2C 1 Putative RNA-binding protein Q99 protein protein human Homo sapiens Human MEAGPSGAAAGAYLPPLQQVFQAPRRPGIGTVGKPIKLLANYFEVDIPKIDVYHYEVDIKPDKCPRRVNREVVEYMVQHFKPQIFGDRKPVYDGKKNIYTVTALPIGNERVDFEVTIPGEGKDRIFKVSIKWLAIVSWRMLHEALVSGQIPVPLESVQALDVAMRHLASMRYTPVGRSFFSPPEGYYHPLGGGREVWFGFHQSVRPAMWKMMLNIDVSATAFYKAQPVIEFMCEVLDIRNIDEQPKPLTDSQRVRFTKEIKGLKVEVTHCGQMKRKYRVCNVTRRPASHQTFPLQLESGQTVECTVAQYFKQKYNLQLKYPHLPCLQVGQEQKHTYLPLEVCNIVAGQRCIKKLTDNQTSTMIKATARSAPDRQEEISRLMKNASYNLDPYIQEFGIKVKDDMTEVTGRVLPAPILQYGGRNRAIATPNQGVWDMRGKQFYNGIEIKVWAIACFAPQKQCREEVLKNFTDQLRKISKDAGMPIQGQPCFCKYAQGADSVEPMFRHLKNTYSGLQLIIVILPGKTPVYAEVKRVGDTLLGMATQCVQVKNVVKTSPQTLSNLCLKINVKLGGINNILVPHQRSAVFQQPVIFLGADVTHPPAGDGKKPSITAVVGSMDAHPSRYCATVRVQRPRQEIIEDLSYMVRELLIQFYKSTRFKPTRIIFYRDGVPEGQLPQILHYELLAIRDACIKLEKDYQPGITYIVVQKRHHTRLFCADKNERIGKSGNIPAGTTVDTNITHPFEFDFYLCSHAGIQGTSRPSHYYVLWDDNRFTADELQILTYQLCHTYVRCTRSVSIPAPAYYARLVAFRARYHLVDKEHDSGEGSHISGQSNGRDPQALAKAVQVHQDTLRTMYFA ago3_human Protein argonaute-3 AGO3 Argonaute RISC catalytic component 3 EIF2C3 Eukaryotic translation initiation factor 2C 3 protein protein human Homo sapiens Human MEIGSAGPAGAQPLLMVPRRPGYGTMGKPIKLLANCFQVEIPKIDVYLYEVDIKPDKCPRRVNREVVDSMVQHFKVTIFGDRRPVYDGKRSLYTANPLPVATTGVDLDVTLPGEGGKDRPFKVSIKFVSRVSWHLLHEVLTGRTLPEPLELDKPISTNPVHAVDVVLRHLPSMKYTPVGRSFFSAPEGYDHPLGGGREVWFGFHQSVRPAMWKMMLNIDVSATAFYKAQPVIQFMCEVLDIHNIDEQPRPLTDSHRVKFTKEIKGLKVEVTHCGTMRRKYRVCNVTRRPASHQTFPLQLENGQTVERTVAQYFREKYTLQLKYPHLPCLQVGQEQKHTYLPLEVCNIVAGQRCIKKLTDNQTSTMIKATARSAPDRQEEISRLVRSANYETDPFVQEFQFKVRDEMAHVTGRVLPAPMLQYGGRNRTVATPSHGVWDMRGKQFHTGVEIKMWAIACFATQRQCREEILKGFTDQLRKISKDAGMPIQGQPCFCKYAQGADSVEPMFRHLKNTYSGLQLIIVILPGKTPVYAEVKRVGDTLLGMATQCVQVKNVIKTSPQTLSNLCLKINVKLGGINNILVPHQRPSVFQQPVIFLGADVTHPPAGDGKKPSIAAVVGSMDAHPSRYCATVRVQRPRQEIIQDLASMVRELLIQFYKSTRFKPTRIIFYRDGVSEGQFRQVLYYELLAIREACISLEKDYQPGITYIVVQKRHHTRLFCADRTERVGRSGNIPAGTTVDTDITHPYEFDFYLCSHAGIQGTSRPSHYHVLWDDNCFTADELQLLTYQLCHTYVRCTRSVSIPAPAYYAHLVAFRARYHLVDKEHDSAEGSHVSGQSNGRDPQALAKAVQIHQDTLRTMYFA tnrc6b-ago2 1467525 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false peptide_d3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_e12 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_g2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_g3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Figure 1 Binding hotspots entered here, however several other sites showed weaker binding. tnrc6b-ago2-1 1467525 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q9UPQ9:p.Trp623Ala mutation disrupting mutation disrupting interaction mutation disrupting alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UPQ9:p.Trp623Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Q9UPQ9:p.Trp623Tyr mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Figure 2 When the entire peptide except the central Trp was substituted by alanines or glycines, binding was noticeably reduced. Trp neighbors with small, not bulky side chains are compatible with the Ago2 interaction (Fig. 2F). The aromatic indole ring of Trp is essential for binding. Trp flanking residues identified in Fig. 1E may modulate the interaction but are not necessary for the Ago2 interaction. tnrc6b-ago1-3 1467526 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false trp-623_trp-634 direct binding direct binding region direct binding certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 6A, Supp 3 Regions surrounding residues W623 and W634 in TNRC6B mediate additional, weaker and, likely, nonspecific contacts to Ago2. tnrc6b-ago1-4 1467527 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait Q9UPQ9:p.Gln627_Ile628delinsGlyGly mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus 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 false Q9UPQ9:p.Trp623Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q9UPQ9:p.Trp634Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false undefined mutation mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 6B, 7F. Mutating all tryptophans except of one abolished Ago2 binding. Two tryptophans are required for efficient binding. Moreover, within this binding region, only the first two tryptophans engage in Ago2 binding. Whereas wt, 11- and 12-aa spacers efficiently bound recombinant Ago2, distances of 8 or 9 aa between the two tryptophans strongly reduced Ago2 binding. tnrc6b-ago1-1 1467528 n/a 1467535 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 n/a 1467533 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 n/a 1467534 unspecified role unspecified role bait bait n_terminus 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1467536 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 association association false false false 5 A tnrc6b-ago1-2 1467529 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait cys600 atto 488 atto 488 certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1467529 5C tnrc6b-ago2-3 1467530 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait Q9UPQ9:p.Trp623Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q9UPQ9:p.[Trp623Ala;Trp634Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 6C tnrc6b-ago1-5 1467531 n/a 1467533 unspecified role unspecified role prey prey lys425 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys693 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys425 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys655 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys844 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false n/a 1467534 unspecified role unspecified role bait bait lys648 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys648 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys648 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false lys645 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus 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 false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false lys661 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1467598 1467593 1467599 1467594 1467600 1467595 1467601 1467596 1467605 1467592 physical association physical association false false false 7, Supp 4. tnrc6b-ago2-4 1467532 n/a 1467533 unspecified role unspecified role prey prey n/a 1467534 unspecified role unspecified role bait bait n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false peptide_g2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false peptide_d3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Figure 3, Supp fig 1 In all three peptides, the main contacts to Ago2 are mediated by protons H Eta2 and H lambda 2 positions (of indole ring), suggesting that these protons are buried in the binding pocket of Ago2. Saturation transfer difference (STD) NMR