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/ Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The main body of the text says there is pre-incubation with PKA- yet (Fig 4) there is also a BLACK bar on the results which is "S1444+PKA" - so presumably NOT pre-incubation- (I am ignoring this as results are the same as for pre-incubation (first column -clear bar), if of lower fluorescence) Peptides were phosphorylated by PKA preincubation. As a negative control a PKA inhibitor was used in some preincubations. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-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 Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay tag visualisation tag visualisation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down tag visualisation tag visualisation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. Autophosphorylation of LRRK2 was detected by westerm Blotting using an anti-pT2483 antibody - T2483 being a previously identified LRRK2 autophosphorylation site (one of many) - see Reference 7 (Gloeckner J. Proteome Res 2010. PMID:20108944) The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Parkinson-related LRRK2 mutation R1441C/G/H impairs PKA phosphorylation of LRRK2 and disrupts its interaction with 14-3-3. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 imex curation Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Affinomics - Interactions curated for the Affinomics consortium. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay ms participant Identification by mass spectrometry ms participant Muda K., Bertinetti D., Gesellchen F., Hermann J.S., von Zweydorf F., Geerlof A., Jacob A., Ueffing M., Gloeckner C.J., Herberg F.W. Parkinsons - Interactions investigated in the context of Parkinsons disease Affinomics - Interactions curated for the Affinomics consortium. both MS and MS/MS are mentioned in the Supplement - but it does not say if/when each is used. I have made Supp table 1 into 3 Interactions:- with full length molecule is phosp-ed alone, when both N-terminal deletion construct AND Full length mol produce results , and phoph. using Deletion construct but NOT Full length mol. I have added a comment to each interaction indicate which construct(s) the results are from. (bearing in mind we do not know if the dashes represent negative results or "not tested".) Results were obtained from either the full length LRRK2 Kinase Dead mutant alone, or the full length molecule concurrently with the truncated Delta-967 LRRK2 Kinase Dead mutant (aas 968-2527), or from the deletion construct alone. The authors confirm that "PKA" is a Human protein, ID is UniProt P17612 and it was expressed in E.coli. and purified by affinity purification. herberg@uni-kassel.de Accepted 2014-FEB-25 by ORCHARD Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2013 Only protein-protein interactions imex curation kapca_human cAMP-dependent protein kinase catalytic subunit alpha PRKACA PKACA protein protein human Homo sapiens Human MGNAAAAKKGSEQESVKEFLAKAKEDFLKKWESPAQNTAHLDQFERIKTLGTGSFGRVMLVKHKETGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFLVKLEFSFKDNSNLYMVMEYVPGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDLIYRDLKPENLLIDQQGYIQVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDWWALGVLIYEMAAGYPPFFADQPIQIYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKRFGNLKNGVNDIKNHKWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSINEKCGKEFSEF DIP protein P17612 original sequence version: 171 DIP protein P17612 original sequence version: 175 DIP protein P17612 original sequence version: 178 DIP protein P17612-1 original sequence version: 177 DIP protein P17612 original sequence version: 186 DIP protein P17612 original sequence version: 102 DIP protein P17612-1 original sequence version: 102 BF6D3ECD2614E5AB moes_human Moesin Membrane-organizing extension spike protein MSN protein protein human Homo sapiens Human MPKTISVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWFFGLQYQDTKGFSTWLKLNKKVTAQDVRKESPLLFKFRAKFYPEDVSEELIQDITQRLFFLQVKEGILNDDIYCPPETAVLLASYAVQSKYGDFNKEVHKSGYLAGDKLLPQRVLEQHKLNKDQWEERIQVWHEEHRGMLREDAVLEYLKIAQDLEMYGVNYFSIKNKKGSELWLGVDALGLNIYEQNDRLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILALCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQMERAMLENEKKKREMAEKEKEKIEREKEELMERLKQIEEQTKKAQQELEEQTRRALELEQERKRAQSEAEKLAKERQEAEEAKEALLQASRDQKKTQEQLALEMAELTARISQLEMARQKKESEAVEWQQKAQMVQEDLEKTRAELKTAMSTPHVAEPAENEQDEQDENGAEASADLRADAMAKDRSEEERTTEAEKNERVQKHLKALTSELANARDESKKTANDMIHAENMRLGRDKYKTLRQIRQGNTKQRIDEFESM DIP protein P26038 original sequence version: 168 865A6C5CB14AE586 1433t_human 14-3-3 protein theta YWHAQ 14-3-3 protein tau 14-3-3 protein T-cell Protein HS1 protein protein human Homo sapiens Human MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIDNSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN DIP protein P27348 original sequence version: 164 DIP protein P27348 original sequence version: 173 175534325E9E37C4 1433g_rat 14-3-3 protein gamma Ywhag protein protein rat Rattus norvegicus (Rat) Rat MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN B0D16C6DE1F4455D 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 1433z_human 14-3-3 protein zeta/delta YWHAZ Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MDKNELVQKAKLAEQAERYDDMAACMKSVTEQGAELSNEERNLLSVAYKNVVGARRSSWRVVSSIEQKTEGAEKKQQMAREYREKIETELRDICNDVLSLLEKFLIPNASQAESKVFYLKMKGDYYRYLAEVAAGDDKKGIVDQSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDTQGDEAEAGEGGEN DIP protein P63104 original sequence version: 144 D464DF2286BBFE60 lrrk2-prkaca-1 8 9 enzyme enzyme unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role Q5S007:p.Lys1906Met mutation mutation certain certain sequence position certain certain certain sequence position certain K M Wed May 02 20:13:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys1906met' Tue Jun 30 17:58:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys1906Met' c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-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 Table 1,2. Supp fig 1. Q5S007 thr-833 O-phospho-L-threonine; Q5S007 ser-850 O-phospho-L-serine; Q5S007 ser-858 O-phospho-L-serine; Q5S007 ser-860 O-phospho-L-serine; Q5S007 ser-908 O-phospho-L-serine; Q5S007 ser-910 O-phospho-L-serine; Q5S007 ser-933 O-phospho-L-serine; Q5S007 ser-935 O-phospho-L-serine; Q5S007 ser-954 O-phospho-L-serine; Q5S007 ser-955 O-phospho-L-serine Q5S007 ser-958 O-phospho-L-serine Q5S007 ser-962 O-phospho-L-serine These phosphorylations were found using only Full Length LRRK2. Six serine residues (S910, S955, S971, S973, S976, and S979) also were found in the negative controls (Table S2), indicating that LRRK2 already was phosphorylated to some extent. lrrk2-prkaca-2 8 9 enzyme enzyme unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Lys1906Met mutation mutation certain certain sequence position certain certain certain sequence position certain K M Wed May 02 16:19:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys1906met' Tue Jun 30 15:42:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys1906Met' c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-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 Table 1,2. Supp fig 1. -i Q5S007 ser-971 O-phospho-L-serine; Q5S007 ser-973 O-phospho-L-serine; Q5S007 ser-976 O-phospho-L-serine; Q5S007 ser-1443 O-phospho-L-serine; Q5S007 ser-1444 O-phospho-L-serine These phosphorylations were found using both the Full Length LRRK2 AND the N-terminal LRRK2 deletion construct. Six serine residues (S910, S955, S971, S973, S976, and S979) also were found in the negative controls (Table S2), indicating that LRRK2 already was phosphorylated to some extent. lrrk2-prkaca-3 8 9 enzyme enzyme unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Lys1906Met mutation mutation certain certain sequence position certain certain certain sequence position certain K M Wed May 02 17:47:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys1906met' Tue Jun 30 16:31:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys1906Met' 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 Table 1,2. Supp fig 1. -ii Q5S007 ser-979 O-phospho-L-serine; Q5S007 ser-2166 O-phospho-L-serine; Q5S007 thr-1849 O-phospho-L-threonine These phosphorylations were found only using the N-terminal LRRK2 deletion construct, and not using the Full length molecule. Six serine residues (S910, S955, S971, S973, S976, and S979) also were found in the negative controls (Table S2), indicating that LRRK2 already was phosphorylated to some extent. lrrk2-prkaca-4 5 9 enzyme enzyme unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-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 1A Q5S007 ser-910 O-phospho-L-serine; Q5S007 ser-935 O-phospho-L-serine LRRK2 already showed basal phosphorylation on both S910 and S935, which were increased significantly upon incubation with PKA. P17612 increases binding of Q5S007 to P61981 lrrk2-prkaca-5 2 9 enzyme enzyme unspecified role unspecified role 13 enzyme target enzyme target substrate substrate unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined roc_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Arg1441Cys mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R C Wed May 02 20:06:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441cys' Tue Jun 30 20:03:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Cys' Q5S007:p.Arg1441His mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R H Wed May 02 19:02:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441his' Tue Jun 30 18:56:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441His' Q5S007:p.Ser1444Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain S A Wed May 02 18:24:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 15:42:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' Q5S007:p.Ser1443_Ser1444delinsAlaAla mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain SS AA Thu Jul 07 12:07:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ser1444ala,ser1443ala Fri Jul 22 11:13:07 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala,ser1443ala' Wed May 02 15:10:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser_ser1443-1444ala_ala' Tue Jun 30 20:23:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1443_Ser1444delinsAlaAla' Q5S007:p.Arg1441Gly mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R G Wed May 02 15:31:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441gly' Tue Jun 30 20:15:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Gly' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Figure 2 P10644 inhibits activity of P17612 ywhag-lrrk2-7 4 13 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain required_phosphoser1444 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) 12 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role prey prey c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 physical association physical association Figure 3ii C LRRK2 WT Delta-967, which does not contain S910/S935, was precipitated efficiently by GST–14-3-3 gamma, strongly indicating an additional 14-3-3 docking site in LRRK2. ywhag-lrrk2-8 4 13 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-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 12 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 3B/C LRRK2 WT Delta-967, which does not contain S910/S935, was precipitated efficiently by GST–14-3-3 gamma, strongly indicating an additional 14-3-3 docking site in LRRK2. prkaca-lrrk2 6 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q5S007:p.Ser1443_Ser1444delinsAlaAla mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain SS AA Thu Jul 07 12:18:13 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ser1444ala,ser1443ala Fri Jul 22 11:02:20 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala,ser1443ala' Wed May 02 20:06:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser_ser1443-1444ala_ala' Tue Jun 30 15:44:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1443_Ser1444delinsAlaAla' Q5S007:p.Ser1444Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A Wed May 02 20:05:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 18:06:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' roc_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain required_phosphoser1444 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 ecoli Escherichia coli (strain K12) 12 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3D Binding of 14-3-3 to LRRK2-ROC was detected exclusively to PKA-phosphorylated but not to nonphosphorylated ROC fusion protein, and could be observed only when S1444 was present. lrrk2-ywhag-2 1 12 unspecified role unspecified role neutral component neutral component 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) 13 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n_terminus fluorescein fluorescein label fluorescein n-term range n-terminal range n-term range n-term range n-terminal range n-term range required_phosphoser1444 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 Q5S007:p.Arg1441Gly mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R G Wed May 02 15:08:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441gly' Tue Jun 30 17:58:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Gly' Q5S007:p.Ser1444Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:33:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 18:31:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figure 4iii, Supp figure 2, Table 1. Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhaq-1 1 13 unspecified role unspecified role neutral component neutral component n_terminus fluorescein fluorescein label fluorescein n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q5S007:p.Ser1444Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:35:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 17:35:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' required_phosphoser1444 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 Q5S007:p.Arg1441Gly mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R G Wed May 02 18:41:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441gly' Tue Jun 30 20:03:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Gly' region 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) 11 unspecified role unspecified role neutral component neutral component 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 Figure 4ii, supp figure 2, Table 1 Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhaz-1 1 13 unspecified role unspecified role neutral component neutral component Q5S007:p.Arg1441Gly mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R G Wed May 02 20:01:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441gly' Tue Jun 30 18:04:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Gly' n_terminus fluorescein fluorescein label fluorescein n-term range n-terminal range n-term range n-term range n-terminal range n-term range required_phosphoser1444 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 Q5S007:p.Ser1444Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A Wed May 02 14:47:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 19:44:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' region 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) 14 unspecified role unspecified role neutral component neutral component 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 Figure 4iii, Table 1, Supp fig 2. Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhag-3 1 12 unspecified role unspecified role neutral component neutral component 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) 13 unspecified role unspecified role neutral component neutral component required_phosphoser910 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n_terminus fluorescein fluorescein label fluorescein n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Table 1, figure S2 Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhaz-2 1 13 unspecified role unspecified role neutral component neutral component n_terminus fluorescein fluorescein label fluorescein 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 required_phosphoser910 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) 14 unspecified role unspecified role neutral component neutral component 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 Table 1, figure S2 Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhaq-2 1 13 unspecified role unspecified role neutral component neutral component n_terminus fluorescein fluorescein label fluorescein n-term range n-terminal range n-term range n-term range n-terminal range n-term range required_phosphoser910 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 region 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) 11 unspecified role unspecified role neutral component neutral component 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 Table 1, figure S2 Phosphorylated LRRK2 WT peptide bound to all three 14-3-3 isoforms, indicated by a high FP signal, whereas the nonphosphorylated WT LRRK2 peptide did not bind to any of the 14-3-3 isoforms. lrrk2-ywhag-1 7 13 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain required_phosphoser1444 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 n_terminus biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 12 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 5 lrrk2-ywhaz 7 13 unspecified role unspecified role bait bait required_phosphoser1444 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n_terminus biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) 14 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 5 lrrk2-ywhaq 7 11 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain required_phosphoser1444 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 n_terminus biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figure 5 lrrk2-prkaca-6 2 10 enzyme target enzyme target substrate substrate unspecified role unspecified role region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 13 enzyme enzyme unspecified role unspecified role c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-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 Fig 6A P26038 phosres lrrk2-1 2 13 putative self putative self putative self putative self c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q5S007:p.Ser1444Ala mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:52:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 16:16:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' 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 16:57:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 17:25:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' Q5S007:p.Arg1441His mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain R H Wed May 02 15:02:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441his' Tue Jun 30 15:38:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441His' 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 Fig 6 A Q5S007 phosres lrrk2-1 3 13 putative self putative self putative self putative self Q5S007:p.Ser1444Ala mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:58:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser1444ala' Tue Jun 30 17:17:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1444Ala' 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 15:16:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 19:01:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q5S007:p.Arg1441His mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain R H Wed May 02 20:22:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441his' Tue Jun 30 18:23:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441His' 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 Fig 6 B Q5S007 phosres lrrk2-1 5 13 putative self putative self putative self putative self c_terminus_strepii/flag_tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q5S007:p.Lys1906Met mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K M Wed May 02 20:00:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys1906met' Tue Jun 30 19:57:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys1906Met' 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 Fig 6 C, Supp fig3 Q5S007 thr-2483 O-phospho-L-threonine PKA together with GST–14-3-3 gamma