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/ LRRK2 regulates tau phosphorylation through direct activation of GSK-3β LRRK2 regulates tau phosphorylation through direct activation of GSK-3β FEBS 2013 imex curation Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Alzheimers - Interactions investigated in the context of Alzheimers disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Parkinsons - Interactions investigated in the context of Parkinsons disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp Accepted 2013-NOV-28 by ORCHARD FEBS 2013 Only protein-protein interactions imex curation LRRK2 regulates tau phosphorylation through direct activation of GSK-3β LRRK2 regulates tau phosphorylation through direct activation of GSK-3β FEBS 2013 imex curation Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Alzheimers - Interactions investigated in the context of Alzheimers disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp 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 anti tag western anti tag western blot anti tag western Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Parkinsons - Interactions investigated in the context of Parkinsons disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp Accepted 2013-NOV-28 by ORCHARD FEBS 2013 Only protein-protein interactions imex curation LRRK2 regulates tau phosphorylation through direct activation of GSK-3β LRRK2 regulates tau phosphorylation through direct activation of GSK-3β FEBS 2013 imex curation Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Alzheimers - Interactions investigated in the context of Alzheimers disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions human-sh_sy5y Homo sapiens SH-SY5Y cell line sh_sy5y Human neuroblastoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Parkinsons - Interactions investigated in the context of Parkinsons disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp Accepted 2013-NOV-28 by ORCHARD FEBS 2013 Only protein-protein interactions imex curation LRRK2 regulates tau phosphorylation through direct activation of GSK-3β LRRK2 regulates tau phosphorylation through direct activation of GSK-3β FEBS 2013 imex curation Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Alzheimers - Interactions investigated in the context of Alzheimers disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Parkinsons - Interactions investigated in the context of Parkinsons disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp Accepted 2013-NOV-28 by ORCHARD FEBS 2013 Only protein-protein interactions imex curation LRRK2 regulates tau phosphorylation through direct activation of GSK-3β LRRK2 regulates tau phosphorylation through direct activation of GSK-3β FEBS 2013 imex curation Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Alzheimers - Interactions investigated in the context of Alzheimers disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Kawakami F, Shimada N, Ohta E, Kagiya G, Kawashima R, Maekawa T, Maruyama H, Ichikawa T. Parkinsons - Interactions investigated in the context of Parkinsons disease Dear Checker- there are lots of errors in this hopefully preliminary version of a publication (no - not just any errors I have made here...) - e.g. missing images/legends + their images/legends the wrong way around, wrong suppliers etc... For some of these errors I have obtained the correct info from the authors - I have also emailed FEBS journal with a copy of the authors replies (he seemed to think it was my job to alter all his errors!)- and have informed the author that I have done this.... (so this might mean the final version will have been corrected???) As a result I have NOT added any "cautions", as should the errors be corrected the cautions would be redundant. In Supplementary figure 2 in some instances GSK3Beta is heat treated (at 95degrees for 5 minutes) to inactivate it's enzymatic activity. Supplementary figure 2C uses 2D TLC electrophoresis followed by autoradiography to show that Threonine residues of GSK-3B were predominantly phosphorylated. Domain numbering for LRRK2 has been taken from UniProt, as this info does not appear in this publication. The authors have stated that 'His tagged GSK3Beta was obtained from Sigma, and that the sequence of the "peptide substrate" (as used in Figure 3) was RRRPASVPPSPSLS RHS(pS)HQRR -based on muscle glycogen synthase 1. The authors also supplied experimental details of heat inactivation of GSK3Beta - as used in Supplementary figure 2. kawakami@kitasato-u.ac.jp Accepted 2013-NOV-28 by ORCHARD FEBS 2013 Only protein-protein interactions imex curation p10636-8 Microtubule-associated protein tau Tau-4 MAPT MAPTL MTBT1 TAU Neurofibrillary tangle protein Paired helical filament-tau 2N4R protein protein human Homo sapiens Human MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL DIP protein P10636-8 original sequence version: 206 DIP protein P10636-8 original sequence version: 203 DIP protein P10636-8 original sequence version: 166 DIP protein P10636-8 original sequence version: 218 835A8706D847A8CC gsk3b_human Glycogen synthase kinase-3 beta GSK3B Serine/threonine-protein kinase GSK3B protein protein human Homo sapiens Human MSGRPRTTSFAESCKPVQQPSAFGSMKVSRDKDGSKVTTVVATPGQGPDRPQEVSYTDTKVIGNGSFGVVYQAKLCDSGELVAIKKVLQDKRFKNRELQIMRKLDHCNIVRLRYFFYSSGEKKDEVYLNLVLDYVPETVYRVARHYSRAKQTLPVIYVKLYMYQLFRSLAYIHSFGICHRDIKPQNLLLDPDTAVLKLCDFGSAKQLVRGEPNVSYICSRYYRAPELIFGATDYTSSIDVWSAGCVLAELLLGQPIFPGDSGVDQLVEIIKVLGTPTREQIREMNPNYTEFKFPQIKAHPWTKVFRPRTPPEAIALCSRLLEYTPTARLTPLEACAHSFFDELRDPNVKLPNGRDTPALFNFTTQELSSNPPLATILIPPHARIQAAASTPTNATAASDANTGDRGQTNNAASASASNST DIP protein P49841 original sequence version: 109 DIP protein P49841 original sequence version: 136 DIP protein P49841 original sequence version: 143 DIP protein P49841 original sequence version: 141 DIP protein P49841 original sequence version: 196 DIP protein P49841 original sequence version: 159 DIP protein P49841 original sequence version: 151 4ACC24D00CDBB9C3 gys1_human Glycogen [starch] synthase, muscle GYS1 GYS protein protein human Homo sapiens Human MPLNRTLSMSSLPGLEDWEDEFDLENAVLFEVAWEVANKVGGIYTVLQTKAKVTGDEWGDNYFLVGPYTEQGVRTQVELLEAPTPALKRTLDSMNSKGCKVYFGRWLIEGGPLVVLLDVGASAWALERWKGELWDTCNIGVPWYDREANDAVLFGFLTTWFLGEFLAQSEEKPHVVAHFHEWLAGVGLCLCRARRLPVATIFTTHATLLGRYLCAGAVDFYNNLENFNVDKEAGERQIYHRYCMERAAAHCAHVFTTVSQITAIEAQHLLKRKPDIVTPNGLNVKKFSAMHEFQNLHAQSKARIQEFVRGHFYGHLDFNLDKTLYFFIAGRYEFSNKGADVFLEALARLNYLLRVNGSEQTVVAFFIMPARTNNFNVETLKGQAVRKQLWDTANTVKEKFGRKLYESLLVGSLPDMNKMLDKEDFTMMKRAIFATQRQSFPPVCTHNMLDDSSDPILTTIRRIGLFNSSADRVKVIFHPEFLSSTSPLLPVDYEEFVRGCHLGVFPSYYEPWGYTPAECTVMGIPSISTNLSGFGCFMEEHIADPSAYGIYILDRRFRSLDDSCSQLTSFLYSFCQQSRRQRIIQRNRTERLSDLLDWKYLGRYYMSARHMALSKAFPEHFTYEPNEADAAQGYRYPRPASVPPSPSLSRHSSPHQSEDEEDPRNGPLEEDGERYDEDEEAAKDRRNIRAPEWPRRASCTSSTSGSKRNSVDTATSSSLSTPSEPLSPTSSLGEERN 0E321BBFDEB0BD7F lrrk2_human Leucine-rich repeat serine/threonine-protein kinase 2 LRRK2 PARK8 Dardarin protein protein human Homo sapiens Human MASGSCQGCEEDEETLKKLIVRLNNVQEGKQIETLVQILEDLLVFTYSERASKLFQGKNIHVPLLIVLDSYMRVASVQQVGWSLLCKLIEVCPGTMQSLMGPQDVGNDWEVLGVHQLILKMLTVHNASVNLSVIGLKTLDLLLTSGKITLLILDEESDIFMLIFDAMHSFPANDEVQKLGCKALHVLFERVSEEQLTEFVENKDYMILLSALTNFKDEEEIVLHVLHCLHSLAIPCNNVEVLMSGNVRCYNIVVEAMKAFPMSERIQEVSCCLLHRLTLGNFFNILVLNEVHEFVVKAVQQYPENAALQISALSCLALLTETIFLNQDLEEKNENQENDDEGEEDKLFWLEACYKALTWHRKNKHVQEAACWALNNLLMYQNSLHEKIGDEDGHFPAHREVMLSMLMHSSSKEVFQASANALSTLLEQNVNFRKILLSKGIHLNVLELMQKHIHSPEVAESGCKMLNHLFEGSNTSLDIMAAVVPKILTVMKRHETSLPVQLEALRAILHFIVPGMPEESREDTEFHHKLNMVKKQCFKNDIHKLVLAALNRFIGNPGIQKCGLKVISSIVHFPDALEMLSLEGAMDSVLHTLQMYPDDQEIQCLGLSLIGYLITKKNVFIGTGHLLAKILVSSLYRFKDVAEIQTKGFQTILAILKLSASFSKLLVHHSFDLVIFHQMSSNIMEQKDQQFLNLCCKCFAKVAMDDYLKNVMLERACDQNNSIMVECLLLLGADANQAKEGSSLICQVCEKESSPKLVELLLNSGSREQDVRKALTISIGKGDSQIISLLLRRLALDVANNSICLGGFCIGKVEPSWLGPLFPDKTSNLRKQTNIASTLARMVIRYQMKSAVEEGTASGSDGNFSEDVLSKFDEWTFIPDSSMDSVFAQSDDLDSEGSEGSFLVKKKSNSISVGEFYRDAVLQRCSPNLQRHSNSLGPIFDHEDLLKRKRKILSSDDSLRSSKLQSHMRHSDSISSLASEREYITSLDLSANELRDIDALSQKCCISVHLEHLEKLELHQNALTSFPQQLCETLKSLTHLDLHSNKFTSFPSYLLKMSCIANLDVSRNDIGPSVVLDPTVKCPTLKQFNLSYNQLSFVPENLTDVVEKLEQLILEGNKISGICSPLRLKELKILNLSKNHISSLSENFLEACPKVESFSARMNFLAAMPFLPPSMTILKLSQNKFSCIPEAILNLPHLRSLDMSSNDIQYLPGPAHWKSLNLRELLFSHNQISILDLSEKAYLWSRVEKLHLSHNKLKEIPPEIGCLENLTSLDVSYNLELRSFPNEMGKLSKIWDLPLDELHLNFDFKHIGCKAKDIIRFLQQRLKKAVPYNRMKLMIVGNTGSGKTTLLQQLMKTKKSDLGMQSATVGIDVKDWPIQIRDKRKRDLVLNVWDFAGREEFYSTHPHFMTQRALYLAVYDLSKGQAEVDAMKPWLFNIKARASSSPVILVGTHLDVSDEKQRKACMSKITKELLNKRGFPAIRDYHFVNATEESDALAKLRKTIINESLNFKIRDQLVVGQLIPDCYVELEKIILSERKNVPIEFPVIDRKRLLQLVRENQLQLDENELPHAVHFLNESGVLLHFQDPALQLSDLYFVEPKWLCKIMAQILTVKVEGCPKHPKGIISRRDVEKFLSKKRKFPKNYMSQYFKLLEKFQIALPIGEEYLLVPSSLSDHRPVIELPHCENSEIIIRLYEMPYFPMGFWSRLINRLLEISPYMLSGRERALRPNRMYWRQGIYLNWSPEAYCLVGSEVLDNHPESFLKITVPSCRKGCILLGQVVDHIDSLMEEWFPGLLEIDICGEGETLLKKWALYSFNDGEEHQKILLDDLMKKAEEGDLLVNPDQPRLTIPISQIAPDLILADLPRNIMLNNDELEFEQAPEFLLGDGSFGSVYRAAYEGEEVAVKIFNKHTSLRLLRQELVVLCHLHHPSLISLLAAGIRPRMLVMELASKGSLDRLLQQDKASLTRTLQHRIALHVADGLRYLHSAMIIYRDLKPHNVLLFTLYPNAAIIAKIADYGIAQYCCRMGIKTSEGTPGFRAPEVARGNVIYNQQADVYSFGLLLYDILTTGGRIVEGLKFPNEFDELEIQGKLPDPVKEYGCAPWPMVEKLIKQCLKENPQERPTSAQVFDILNSAELVCLTRRILLPKNVIVECMVATHHNSRNASIWLGCGHTDRGQLSFLDLNTEGYTSEEVADSRILCLALVHLPVEKESWIVSGTQSGTLLVINTEDGKKRHTLEKMTDSVTCLYCNSFSKQSKQKNFLLVGTADGKLAIFEDKTVKLKGAAPLKILNIGNVSTPLMCLSESTNSTERNVMWGGCGTKIFSFSNDFTIQKLIETRTSQLFSYAAFSDSNIITVVVDTALYIAKQNSPVVEVWDKKTEKLCGLIDCVHFLREVMVKENKESKHKMSYSGRVKTLCLQKNTALWIGTGGGHILLLDLSTRRLIRVIYNFCNSVRVMMTAQLGSLKNVMLVLGYNRKNTEGTQKQKEIQSCLTVWDINLPHEVQNLEKHIEVRKELAEKMRRTSVE DIP protein Q5S007 original sequence version: 36 DIP protein Q5S007 original sequence version: 68 DIP protein Q5S007 original sequence version: 64 26142A0CECBBC3F4 lrrk2-gsk3b-1 2 7 unspecified role unspecified role prey prey c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role bait bait c_terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association 2A lrrk2-gsk3b-2 3 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association 2B lrrk2-gsk3b-3 1 7 unspecified role unspecified role prey prey 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 unspecified role unspecified role bait bait Q5S007:p.Gly2019Ser mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain G S Wed May 02 16:07:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 19:04:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' kinase_domain sufficient to bind sufficient binding region sufficient to bind range range range range 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 2 C/D, Fig 4A (B) gsk3b-lrrk2-4 5 9 stimulator stimulator ancillary ancillary n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q5S007:p.Gly2019Ser mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain G S Wed May 02 19:31:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 18:22:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' ecoli Escherichia coli (strain K12) 7 enzyme enzyme neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 3 E/F, 4A (B) the image legend states that figs 3(c)/d use tau - but this does not make sense as the peptide substrates are also present...so not annotated. Q5S007 increases activity of P49841 gsk3b-lrrk2-7 5 9 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q5S007:p.Asp1994Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:06:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1994ala' Tue Jun 30 18:26:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1994Ala' ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation Supp 2i Using kinase-dead LRRK2 (D1994A) instead of Wild-type, the interaction was abolished. (heat inactivated GSK3-beta was used). gsk3b-1 5 7 putative self putative self putative self putative self 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation true Supp 2ii gsk3b-lrrk2-8 4 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction Supp figure 3 LRRK2 does not enhance the binding affinity of GSK-3B with tau. gsk3b-gys1-2 5 8 enzyme target enzyme target substrate substrate neutral component neutral component c_terminal_his,gln,arg,arg sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ser652 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 7 enzyme enzyme neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 stimulator stimulator ancillary ancillary 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 3 A/B LRRK2 -Wild type or Kinase Dead (D1994A) LRRK2 directly increases the phosphorylation activity of GSK-3Beta for exogenous substrate, and this stimulatory effect of LRRK2 does not involve its kinase activity. lrrk2-1 5 9 putative self putative self putative self putative self Q5S007:p.Asp1994Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 19:03:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1994ala' Tue Jun 30 18:00:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1994Ala' n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation true Supp 2iii Using kinase-dead LRRK2 (D1994A) instead of Wild-type, this interaction was abolished. (also "heat" GSK3-beta was present). gsk3b-lrrk2-9 4 7 unspecified role unspecified role bait bait 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association Supp figure 3ii LRRK2 does not enhance the binding affinity of GSK-3B with tau. gsk3b-gys1-3 5 7 enzyme enzyme neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 8 enzyme target enzyme target substrate substrate neutral component neutral component c_terminal_his,gln,arg,arg sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ser652 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation 3 A/B gsk3b-lrrk2-10 5 6 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme enzyme neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation 3E/F, 4A (B) the image legend states that figs 3(c)/d use tau - but this does not make sense as the peptide substrates are also present...so not annotated. Q5S007 increases activity of P49841