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/ Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... Saturation transfer difference (STD) NMR niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Structural features of Argonaute-GW182 protein interactions. Structural features of Argonaute-GW182 protein interactions. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Pfaff J., Hennig J., Herzog F., Aebersold R., Sattler M., Niessing D., Meister G. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. 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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. 13-Jan-2014: Contacted by IntAct-Help. Only protein-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. I dont think fig 5B - DLS- shows binding as the bottom image with both interactors peaks look to be much the the same as the two higher ones (- so not entered), but the text says "However, when measured together, the peak is slightly shifted to higher molecular weights (Fig. 5B, Bottom), indicating that the complex is not aggregated unspecifically and can be used for further analysis." -dont quite know what they are getting at here... Cross linker was:- disuccinimidyl suberate niessing@helmholtz-muenchen.de,gunter.meister@vkl.uni-regensburg.de. Accepted 2013-DEC-19 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation 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 DIP protein Q9H9G7 original sequence version: 81 6FF1277995E5322E 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 DIP protein Q9UL18 original sequence version: 46 DIP protein Q9UL18 original sequence version: 57 DIP protein Q9UL18 original sequence version: 80 DIP protein Q9UL18 original sequence version: 99 1DBB524AE7CBAF66 tnr6b_human Trinucleotide repeat-containing gene 6B protein TNRC6B KIAA1093 protein protein human Homo sapiens Human MREKEQEREEQLMEDKKRKKEDKKKKEATQKVTEQKTKVPEVTKPSLSQPTAASPIGSSPSPPVNGGNNAKRVAVPNGQPPSAARYMPREVPPRFRCQQDHKVLLKRGQPPPPSCMLLGGGAGPPPCTAPGANPNNAQVTGALLQSESGTAPDSTLGGAAASNYANSTWGSGASSNNGTSPNPIHIWDKVIVDGSDMEEWPCIASKDTESSSENTTDNNSASNPGSEKSTLPGSTTSNKGKGSQCQSASSGNECNLGVWKSDPKAKSVQSSNSTTENNNGLGNWRNVSGQDRIGPGSGFSNFNPNSNPSAWPALVQEGTSRKGALETDNSNSSAQVSTVGQTSREQQSKMENAGVNFVVSGREQAQIHNTDGPKNGNTNSLNLSSPNPMENKGMPFGMGLGNTSRSTDAPSQSTGDRKTGSVGSWGAARGPSGTDTVSGQSNSGNNGNNGKEREDSWKGASVQKSTGSKNDSWDNNNRSTGGSWNFGPQDSNDNKWGEGNKMTSGVSQGEWKQPTGSDELKIGEWSGPNQPNSSTGAWDNQKGHPLPENQGNAQAPCWGRSSSSTGSEVGGQSTGSNHKAGSSDSHNSGRRSYRPTHPDCQAVLQTLLSRTDLDPRVLSNTGWGQTQIKQDTVWDIEEVPRPEGKSDKGTEGWESAATQTKNSGGWGDAPSQSNQMKSGWGELSASTEWKDPKNTGGWNDYKNNNSSNWGGGRPDEKTPSSWNENPSKDQGWGGGRQPNQGWSSGKNGWGEEVDQTKNSNWESSASKPVSGWGEGGQNEIGTWGNGGNASLASKGGWEDCKRSPAWNETGRQPNSWNKQHQQQQPPQQPPPPQPEASGSWGGPPPPPPGNVRPSNSSWSSGPQPATPKDEEPSGWEEPSPQSISRKMDIDDGTSAWGDPNSYNYKNVNLWDKNSQGGPAPREPNLPTPMTSKSASVWSKSTPPAPDNGTSAWGEPNESSPGWGEMDDTGASTTGWGNTPANAPNAMKPNSKSMQDGWGESDGPVTGARHPSWEEEEDGGVWNTTGSQGSASSHNSASWGQGGKKQMKCSLKGGNNDSWMNPLAKQFSNMGLLSQTEDNPSSKMDLSVGSLSDKKFDVDKRAMNLGDFNDIMRKDRSGFRPPNSKDMGTTDSGPYFEKLTLPFSNQDGCLGDEAPCSPFSPSPSYKLSPSGSTLPNVSLGAIGTGLNPQNFAARQGGSHGLFGNSTAQSRGLHTPVQPLNSSPSLRAQVPPQFISPQVSASMLKQFPNSGLSPGLFNVGPQLSPQQIAMLSQLPQIPQFQLACQLLLQQQQQQQLLQNQRKISQAVRQQQEQQLARMVSALQQQQQQQQRQPGMKHSPSHPVGPKPHLDNMVPNALNVGLPDLQTKGPIPGYGSGFSSGGMDYGMVGGKEAGTESRFKQWTSMMEGLPSVATQEANMHKNGAIVAPGKTRGGSPYNQFDIIPGDTLGGHTGPAGDSWLPAKSPPTNKIGSKSSNASWPPEFQPGVPWKGIQNIDPESDPYVTPGSVLGGTATSPIVDTDHQLLRDNTTGSNSSLNTSLPSPGAWPYSASDNSFTNVHSTSAKFPDYKSTWSPDPIGHNPTHLSNKMWKNHISSRNTTPLPRPPPGLTNPKPSSPWSSTAPRSVRGWGTQDSRLASASTWSDGGSVRPSYWLVLHNLTPQIDGSTLRTICMQHGPLLTFHLNLTQGTALIRYSTKQEAAKAQTALHMCVLGNTTILAEFATDDEVSRFLAQAQPPTPAATPSAPAAGWQSLETGQNQSDPVGPALNLFGGSTGLGQWSSSAGGSSGADLAGASLWGPPNYSSSLWGVPTVEDPHRMGSPAPLLPGDLLGGGSDSI DIP protein Q9UPQ9 original sequence version: 73 E982B786C97C75D0 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 DIP protein Q9UKV8 original sequence version: 1(def) DIP protein Q9UKV8 original sequence version: 44 DIP protein Q9UKV8 original sequence version: 60 DIP protein Q9UKV8 original sequence version: 62 DIP protein Q9UKV8 original sequence version: 89 DIP protein Q9UKV8 original sequence version: 85 DIP protein Q9UKV8 original sequence version: 87 DIP protein Q9UKV8 original sequence version: 99 DIP protein Q9UKV8 original sequence version: 117 DIP protein Q9UKV8-1 original sequence version: 117 DIP protein Q9UKV8 original sequence version: 145 DIP protein Q9UKV8 original sequence version: 108 5C8552C43FC81345 tnrc6b-ago2 4 12 unspecified role unspecified role prey prey 11 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 peptide_d3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_e12 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_g2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_g3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figure 1 Binding hotspots entered here, however several other sites showed weaker binding. tnrc6b-ago2-1 4 12 unspecified role unspecified role prey prey 11 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 Q9UPQ9:p.Trp623Ala mutation disrupting mutation disrupting interaction mutation disrupting alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain W A Wed May 02 17:07:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623ala' Tue Jun 30 19:22:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp623Ala' peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9UPQ9:p.Trp623Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W F Wed May 02 18:14:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623phe' Tue Jun 30 16:15:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp623Phe' Q9UPQ9:p.Trp623Tyr mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W Y Wed May 02 16:00:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623tyr' Tue Jun 30 16:20:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp623Tyr' chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 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-ago1-1 5 10 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 12 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 11 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 9 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 5 A tnrc6b-ago1-2 2 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait cys600 atto 488 atto 488 certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 5C tnrc6b-ago1-3 6 12 unspecified role unspecified role prey prey 11 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 trp-623_trp-634 direct binding direct binding region direct binding certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 6A, Supp 3 W623 +634 entered as 'Direct binding Region'- OLS says to use with NMR(XRay) - "The defined region of protein which makes physical contact with the interacting partner." Regions surrounding residues W623 and W634 in TNRC6B mediate additional, weaker and, likely, nonspecific contacts to Ago2. tnrc6b-ago1-4 7 12 unspecified role unspecified role prey prey 11 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 QI GG Thu Jul 07 12:18:18 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was gln627_628ile Fri Jul 22 11:07:04 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'gln627gly,ile62gly' Wed May 02 19:18:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln_ile627-628gly_gly' Tue Jun 30 19:46:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln627_Ile628delinsGlyGly' 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 Q9UPQ9:p.Trp623Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W A Wed May 02 20:15:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623ala' Tue Jun 30 16:09:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp623Ala' Q9UPQ9:p.Trp634Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W A Wed May 02 17:21:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp634ala' Tue Jun 30 15:47:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp634Ala' undefined mutation mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:20:51 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'ile628' ecoli Escherichia coli (strain K12) direct interaction direct interaction 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-ago2-3 3 12 unspecified role unspecified role prey prey 11 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 W A Wed May 02 17:59:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623ala' Tue Jun 30 18:11:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp623Ala' Q9UPQ9:p.[Trp623Ala;Trp634Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain W A Wed May 02 15:10:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp623ala,trp634ala' Tue Jun 30 16:31:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp623Ala;Trp634Ala]' 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 6C tnrc6b-ago1-5 8 12 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 lys693 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain lys425 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain lys655 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain lys844 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain 11 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 lys648 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain lys648 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain lys645 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 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 lys661 xlnk-res-res crosslinked residues XLNK-Res-Res certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 81 68 74 69 75 70 71 76 72 77 physical association physical association 7, Supp 4. tnrc6b-ago2-4 1 12 unspecified role unspecified role prey prey 11 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 peptide_g2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_e11 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain peptide_d3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 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