IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ lobbestael-2013-7 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-3 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-1 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. in vitro In vitro protein phosphatase assay protein phosphatase assay western blot western blot Immuno blot Immuno blot Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. LRRK2 was metabolically labelled with 32P in HEK-293T cells, purified by IP, and incubated with phosphatases at a limiting concentration. Dephosphorylation of LRRK2 was measured using phosphosite-specific anti-LRRK2 antibodies. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-2 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. in vitro In vitro protein phosphatase assay protein phosphatase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. LRRK2 was metabolically labelled with 32P in HEK-293T cells, purified by IP, and incubated with phosphatases at a limiting concentration. After autoradiography on the gel the presence of LRRK2 was confirmed by Western Blot. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-4 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-5 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. lobbestael-2013-6 Identification of protein phosphatase 1 as a regulator of the LRRK2 phosphorylation cycle. human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lobbestael E., Zhao J., Rudenko I.N., Beylina A., Gao F., Wetter J., Beullens M., Bollen M., Cookson M.R., Baekelandt V., Nichols R.J., Taymans J.M. Affinomics - Interactions curated for the Affinomics consortium. Results from 14-3-3 overlay assays cannot be included as the identity of the 14-3-3 protein is not given in this publication nor in (methods section) reference 6 or 9. HSP90 cannot be included as an interactor as it's identity is not given, similarly for MBP. jnichols@parkinsonsinstitute.org,jean-marc.taymans@med.kuleuven.be Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. ppb_ecoli Alkaline phosphatase phoA b0383 JW0374 protein protein ecoli Escherichia coli (strain K12) MKQSTIALALLPLLFTPVTKARTPEMPVLENRAAQGDITAPGGARRLTGDQTAALRDSLSDKPAKNIILLIGDGMGDSEITAARNYAEGAGGFFKGIDALPLTGQYTHYALNKKTGKPDYVTDSAASATAWSTGVKTYNGALGVDIHEKDHPTILEMAKAAGLATGNVSTAELQDATPAALVAHVTSRKCYGPSATSEKCPGNALEKGGKGSITEQLLNARADVTLGGGAKTFAETATAGEWQGKTLREQAQARGYQLVSDAASLNSVTEANQQKPLLGLFADGNMPVRWLGPKATYHGNIDKPAVTCTPNPQRNDSVPTLAQMTDKAIELLSKNEKGFFLQVEGASIDKQDHAANPCGQIGETVDLDEAVQRALEFAKKEGNTLVIVTADHAHASQIVAPDTKAPGLTQALNTKDGAVMVMSYGNSEEDSQEHTGSQLRIAAYGPHAANVVGLTDQTDLFYTMKAALGLK 4-nitrophenyl dihydrogen phosphate 4-nitrophenyl dihydrogen phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ppbi_bovin Intestinal-type alkaline phosphatase ALPI protein protein bovin Bos taurus (Bovine) Bovine MQGACVLLLLGLHLQLSLGLVPVEEEDPAFWNRQAAQALDVAKKLQPIQTAAKNVILFLGDGMGVPTVTATRILKGQMNGKLGPETPLAMDQFPYVALSKTYNVDRQVPDSAGTATAYLCGVKGNYRTIGVSAAARYNQCKTTRGNEVTSVMNRAKKAGKSVGVVTTTRVQHASPAGAYAHTVNRNWYSDADLPADAQMNGCQDIAAQLVNNMDIDVILGGGRKYMFPVGTPDPEYPDDASVNGVRKRKQNLVQAWQAKHQGAQYVWNRTALLQAADDSSVTHLMGLFEPADMKYNVQQDHTKDPTLQEMTEVALRVVSRNPRGFYLFVEGGRIDHGHHDDKAYMALTEAGMFDNAIAKANELTSELDTLILVTADHSHVFSFGGYTLRGTSIFGLAPSKALDSKSYTSILYGNGPGYALGGGSRPDVNDSTSEDPSYQQQAAVPQASETHGGEDVAVFARGPQAHLVHGVEEETFVAHIMAFAGCVEPYTDCNLPAPTTATSIPDAAHLAASPPPLALLAGAMLLLLAPTLY pp1a_human Serine/threonine-protein phosphatase PP1-alpha catalytic subunit PPP1CA PPP1A protein protein human Homo sapiens Human MSDSEKLNLDSIIGRLLEVQGSRPGKNVQLTENEIRGLCLKSREIFLSQPILLELEAPLKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDQGLLCDLLWSDPDKDVQGWGENDRGVSFTFGAEVVAKFLHKHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGAMMSVDETLMCSFQILKPADKNKGKYGQFSGLNPGGRPITPPRNSAKAKK ppm1a_human Protein phosphatase 1A Protein phosphatase 2C isoform alpha PPM1A PPPM1A Protein phosphatase IA protein protein human Homo sapiens Human MGAFLDKPKMEKHNAQGQGNGLRYGLSSMQGWRVEMEDAHTAVIGLPSGLESWSFFAVYDGHAGSQVAKYCCEHLLDHITNNQDFKGSAGAPSVENVKNGIRTGFLEIDEHMRVMSEKKHGADRSGSTAVGVLISPQHTYFINCGDSRGLLCRNRKVHFFTQDHKPSNPLEKERIQNAGGSVMIQRVNGSLAVSRALGDFDYKCVHGKGPTEQLVSPEPEVHDIERSEEDDQFIILACDGIWDVMGNEELCDFVRSRLEVTDDLEKVCNEVVDTCLYKGSRDNMSVILICFPNAPKVSPEAVKKEAELDKYLECRVEEIIKKQGEGVPDLVHVMRTLASENIPSLPPGGELASKRNVIEAVYNRLNPYKNDDTDSTSTDDMW lrrk2_human Leucine-rich repeat serine/threonine-protein kinase 2 LRRK2 PARK8 Dardarin protein protein human Homo sapiens Human MASGSCQGCEEDEETLKKLIVRLNNVQEGKQIETLVQILEDLLVFTYSERASKLFQGKNIHVPLLIVLDSYMRVASVQQVGWSLLCKLIEVCPGTMQSLMGPQDVGNDWEVLGVHQLILKMLTVHNASVNLSVIGLKTLDLLLTSGKITLLILDEESDIFMLIFDAMHSFPANDEVQKLGCKALHVLFERVSEEQLTEFVENKDYMILLSALTNFKDEEEIVLHVLHCLHSLAIPCNNVEVLMSGNVRCYNIVVEAMKAFPMSERIQEVSCCLLHRLTLGNFFNILVLNEVHEFVVKAVQQYPENAALQISALSCLALLTETIFLNQDLEEKNENQENDDEGEEDKLFWLEACYKALTWHRKNKHVQEAACWALNNLLMYQNSLHEKIGDEDGHFPAHREVMLSMLMHSSSKEVFQASANALSTLLEQNVNFRKILLSKGIHLNVLELMQKHIHSPEVAESGCKMLNHLFEGSNTSLDIMAAVVPKILTVMKRHETSLPVQLEALRAILHFIVPGMPEESREDTEFHHKLNMVKKQCFKNDIHKLVLAALNRFIGNPGIQKCGLKVISSIVHFPDALEMLSLEGAMDSVLHTLQMYPDDQEIQCLGLSLIGYLITKKNVFIGTGHLLAKILVSSLYRFKDVAEIQTKGFQTILAILKLSASFSKLLVHHSFDLVIFHQMSSNIMEQKDQQFLNLCCKCFAKVAMDDYLKNVMLERACDQNNSIMVECLLLLGADANQAKEGSSLICQVCEKESSPKLVELLLNSGSREQDVRKALTISIGKGDSQIISLLLRRLALDVANNSICLGGFCIGKVEPSWLGPLFPDKTSNLRKQTNIASTLARMVIRYQMKSAVEEGTASGSDGNFSEDVLSKFDEWTFIPDSSMDSVFAQSDDLDSEGSEGSFLVKKKSNSISVGEFYRDAVLQRCSPNLQRHSNSLGPIFDHEDLLKRKRKILSSDDSLRSSKLQSHMRHSDSISSLASEREYITSLDLSANELRDIDALSQKCCISVHLEHLEKLELHQNALTSFPQQLCETLKSLTHLDLHSNKFTSFPSYLLKMSCIANLDVSRNDIGPSVVLDPTVKCPTLKQFNLSYNQLSFVPENLTDVVEKLEQLILEGNKISGICSPLRLKELKILNLSKNHISSLSENFLEACPKVESFSARMNFLAAMPFLPPSMTILKLSQNKFSCIPEAILNLPHLRSLDMSSNDIQYLPGPAHWKSLNLRELLFSHNQISILDLSEKAYLWSRVEKLHLSHNKLKEIPPEIGCLENLTSLDVSYNLELRSFPNEMGKLSKIWDLPLDELHLNFDFKHIGCKAKDIIRFLQQRLKKAVPYNRMKLMIVGNTGSGKTTLLQQLMKTKKSDLGMQSATVGIDVKDWPIQIRDKRKRDLVLNVWDFAGREEFYSTHPHFMTQRALYLAVYDLSKGQAEVDAMKPWLFNIKARASSSPVILVGTHLDVSDEKQRKACMSKITKELLNKRGFPAIRDYHFVNATEESDALAKLRKTIINESLNFKIRDQLVVGQLIPDCYVELEKIILSERKNVPIEFPVIDRKRLLQLVRENQLQLDENELPHAVHFLNESGVLLHFQDPALQLSDLYFVEPKWLCKIMAQILTVKVEGCPKHPKGIISRRDVEKFLSKKRKFPKNYMSQYFKLLEKFQIALPIGEEYLLVPSSLSDHRPVIELPHCENSEIIIRLYEMPYFPMGFWSRLINRLLEISPYMLSGRERALRPNRMYWRQGIYLNWSPEAYCLVGSEVLDNHPESFLKITVPSCRKGCILLGQVVDHIDSLMEEWFPGLLEIDICGEGETLLKKWALYSFNDGEEHQKILLDDLMKKAEEGDLLVNPDQPRLTIPISQIAPDLILADLPRNIMLNNDELEFEQAPEFLLGDGSFGSVYRAAYEGEEVAVKIFNKHTSLRLLRQELVVLCHLHHPSLISLLAAGIRPRMLVMELASKGSLDRLLQQDKASLTRTLQHRIALHVADGLRYLHSAMIIYRDLKPHNVLLFTLYPNAAIIAKIADYGIAQYCCRMGIKTSEGTPGFRAPEVARGNVIYNQQADVYSFGLLLYDILTTGGRIVEGLKFPNEFDELEIQGKLPDPVKEYGCAPWPMVEKLIKQCLKENPQERPTSAQVFDILNSAELVCLTRRILLPKNVIVECMVATHHNSRNASIWLGCGHTDRGQLSFLDLNTEGYTSEEVADSRILCLALVHLPVEKESWIVSGTQSGTLLVINTEDGKKRHTLEKMTDSVTCLYCNSFSKQSKQKNFLLVGTADGKLAIFEDKTVKLKGAAPLKILNIGNVSTPLMCLSESTNSTERNVMWGGCGTKIFSFSNDFTIQKLIETRTSQLFSYAAFSDSNIITVVVDTALYIAKQNSPVVEVWDKKTEKLCGLIDCVHFLREVMVKENKESKHKMSYSGRVKTLCLQKNTALWIGTGGGHILLLDLSTRRLIRVIYNFCNSVRVMMTAQLGSLKNVMLVLGYNRKNTEGTQKQKEIQSCLTVWDINLPHEVQNLEKHIEVRKELAEKMRRTSVE pp1b_human Serine/threonine-protein phosphatase PP1-beta catalytic subunit PPP1CB protein protein human Homo sapiens Human MADGELNVDSLITRLLEVRGCRPGKIVQMTEAEVRGLCIKSREIFLSQPILLELEAPLKICGDIHGQYTDLLRLFEYGGFPPEANYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRFNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDTGLLCDLLWSDPDKDVQGWGENDRGVSFTFGADVVSKFLNRHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGGMMSVDETLMCSFQILKPSEKKAKYQYGGLNSGRPVTPPRTANPPKKR pp1g_human Serine/threonine-protein phosphatase PP1-gamma catalytic subunit PPP1CC Protein phosphatase 1C catalytic subunit protein protein human Homo sapiens Human MADLDKLNIDSIIQRLLEVRGSKPGKNVQLQENEIRGLCLKSREIFLSQPILLELEAPLKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFLLRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIVDEKIFCCHGGLSPDLQSMEQIRRIMRPTDVPDQGLLCDLLWSDPDKDVLGWGENDRGVSFTFGAEVVAKFLHKHDLDLICRAHQVVEDGYEFFAKRQLVTLFSAPNYCGEFDNAGAMMSVDETLMCSFQILKPAEKKKPNATRPVTPPRGMITKQAKK 4_nitrophenyl_dihydrogen_phosphate-phoa 1564352 n/a 1564359 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1564360 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) dephosphorylation dephosphorylation reaction dephosphorylation false false false Supp fig 3B 4_nitrophenyl_dihydrogen_phosphate-phoa-1 1564352 n/a 1564361 enzyme enzyme neutral component neutral component n/a 1564360 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) dephosphorylation dephosphorylation reaction dephosphorylation false false false Supp fig 3B 4_nitrophenyl_dihydrogen_phosphate-alpi 1564352 n/a 1564360 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1564362 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Supp fig 3B 4_nitrophenyl_dihydrogen_phosphate-ppp1ca 1564352 n/a 1564363 enzyme enzyme neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1564360 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) dephosphorylation dephosphorylation reaction dephosphorylation false false false Supp fig 3B lrrk2-ppp1ca-4 1564353 n/a 1564362 unspecified role unspecified role neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1564364 unspecified role unspecified role neutral component neutral component n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false colocalization colocalization false false false Figure 2A/C lrrk2-ppp1ca-5 1564353 n/a 1564364 unspecified role unspecified role neutral component neutral component n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1564365 unspecified role unspecified role neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false colocalization colocalization false false false Figure 2A-2 lrrk2-ppp1ca-6 1564353 n/a 1564366 unspecified role unspecified role neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1564364 unspecified role unspecified role neutral component neutral component n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false colocalization colocalization false false false Figure 2A-3 LRRK2 co-localized with the (small) fraction of PP1-gamma present in the cytoplasm lrrk2-ppp1ca 1564354 n/a 1564362 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1564364 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ser910 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 false ser935 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 false ser955 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 false ser973 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 false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. dephosphorylation dephosphorylation reaction dephosphorylation false false false Figure 1 lrrk2-ppp1ca-2 1564354 n/a 1564364 enzyme target enzyme target substrate substrate neutral component neutral component ser973 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 false ser935 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 false ser910 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 false ser955 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 false n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. n/a 1564359 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Figure 1 positive control lrrk2-ppp1ca-1 1564355 n/a 1564364 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. n/a 1564362 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Figure 1 lrrk2-ppp1ca-3 1564355 n/a 1564364 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. n/a 1564359 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Figure 1 positive control lrrk2-ppp1ca-8 1564356 n/a 1564364 unspecified role unspecified role bait bait n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ser935 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 false Q5S007:p.Asn1437His mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q5S007:p.Arg1441Gly mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q5S007:p.Tyr1699Cys mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q5S007:p.[Ser910Ala;Ser935Ala;Ser955Ala;Ser973Ala] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1564362 unspecified role unspecified role prey prey n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Figure 2B/D LRRK2-IN1 - a selective LRRK2 inhibitor. [it induces dose-dependent inhibition of Ser910 and Ser935 phosphorylation accompanied by loss of 14-3-3 binding to both wild-type LRRK2 and LRRK2[G2019S] ] Calyculin A -an inhibitor of PP1 and PP2A Addition of LRRK2-IN1 to (WT LRRK2) resulted in ser935 dephosphorylation of LRRK2 and loss of 14-3-3 binding. LRRK2 mutants (which have reduced basal phosphorylation levels) showed enhanced binding to PP1A. A HSP90 interaction with LRRK2 was observed that was also enhanced with LRRK2 (aggregation-prone) mutations. lrrk2-ppp1ca-10 1564357 n/a 1564364 unspecified role unspecified role neutral component neutral component n_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1564362 unspecified role unspecified role neutral component neutral component n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2C LRRK2-IN1 - a selective LRRK2 inhibitor. [it induces dose-dependent inhibition of Ser910 and Ser935 phosphorylation accompanied by loss of 14-3-3 binding to both wild-type LRRK2 and LRRK2[G2019S] ] Calyculin A -an inhibitor of PP1 and PP2A lrrk2-ppp1ca-11 1564358 n/a 1564362 unspecified role unspecified role prey prey n/a 1564364 unspecified role unspecified role bait bait ser935 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 false physical association physical association false false false Figure 2E LRRK2-IN1 - a selective LRRK2 inhibitor. [it induces dose-dependent inhibition of Ser910 and Ser935 phosphorylation accompanied by loss of 14-3-3 binding to both wild-type LRRK2 and LRRK2[G2019S] ] Calyculin A -an inhibitor of PP1 and PP2A Addition of LRRK2-IN1 to (WT LRRK2) resulted in ser935 dephosphorylation of LRRK2.