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/ ray-2014-1 The Parkinson's disease-linked LRRK2 mutation I2020T stabilizes an active state conformation leading to increased kinase activity. in vitro In vitro protein kinase assay protein kinase assay tag visualisation tag visualisation Ray S., Bender S., Kang S., Lin R., Glicksman M.A., Liu M. Parkinsons - Interactions investigated in the context of Parkinsons disease Invitrogen supplied LRRK2 (amino acids 970–2527) and LRRK2 mutation I2020T (also amino acids 970–2527) expressed in a baculovirus system. Both proteins are GST tagged, the mutant with an N-terminal tag, but no tag location is given for the WT protein. J. Biol. Chem. (0021-9258) 2014 imex curation Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Only protein-protein interactions moes_human Moesin Membrane-organizing extension spike protein MSN protein protein human Homo sapiens Human MPKTISVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWFFGLQYQDTKGFSTWLKLNKKVTAQDVRKESPLLFKFRAKFYPEDVSEELIQDITQRLFFLQVKEGILNDDIYCPPETAVLLASYAVQSKYGDFNKEVHKSGYLAGDKLLPQRVLEQHKLNKDQWEERIQVWHEEHRGMLREDAVLEYLKIAQDLEMYGVNYFSIKNKKGSELWLGVDALGLNIYEQNDRLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILALCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQMERAMLENEKKKREMAEKEKEKIEREKEELMERLKQIEEQTKKAQQELEEQTRRALELEQERKRAQSEAEKLAKERQEAEEAKEALLQASRDQKKTQEQLALEMAELTARISQLEMARQKKESEAVEWQQKAQMVQEDLEKTRAELKTAMSTPHVAEPAENEQDEQDENGAEASADLRADAMAKDRSEEERTTEAEKNERVQKHLKALTSELANARDESKKTANDMIHAENMRLGRDKYKTLRQIRQGNTKQRIDEFESM lrrk2_human Leucine-rich repeat serine/threonine-protein kinase 2 LRRK2 PARK8 Dardarin protein protein human Homo sapiens Human MASGSCQGCEEDEETLKKLIVRLNNVQEGKQIETLVQILEDLLVFTYSERASKLFQGKNIHVPLLIVLDSYMRVASVQQVGWSLLCKLIEVCPGTMQSLMGPQDVGNDWEVLGVHQLILKMLTVHNASVNLSVIGLKTLDLLLTSGKITLLILDEESDIFMLIFDAMHSFPANDEVQKLGCKALHVLFERVSEEQLTEFVENKDYMILLSALTNFKDEEEIVLHVLHCLHSLAIPCNNVEVLMSGNVRCYNIVVEAMKAFPMSERIQEVSCCLLHRLTLGNFFNILVLNEVHEFVVKAVQQYPENAALQISALSCLALLTETIFLNQDLEEKNENQENDDEGEEDKLFWLEACYKALTWHRKNKHVQEAACWALNNLLMYQNSLHEKIGDEDGHFPAHREVMLSMLMHSSSKEVFQASANALSTLLEQNVNFRKILLSKGIHLNVLELMQKHIHSPEVAESGCKMLNHLFEGSNTSLDIMAAVVPKILTVMKRHETSLPVQLEALRAILHFIVPGMPEESREDTEFHHKLNMVKKQCFKNDIHKLVLAALNRFIGNPGIQKCGLKVISSIVHFPDALEMLSLEGAMDSVLHTLQMYPDDQEIQCLGLSLIGYLITKKNVFIGTGHLLAKILVSSLYRFKDVAEIQTKGFQTILAILKLSASFSKLLVHHSFDLVIFHQMSSNIMEQKDQQFLNLCCKCFAKVAMDDYLKNVMLERACDQNNSIMVECLLLLGADANQAKEGSSLICQVCEKESSPKLVELLLNSGSREQDVRKALTISIGKGDSQIISLLLRRLALDVANNSICLGGFCIGKVEPSWLGPLFPDKTSNLRKQTNIASTLARMVIRYQMKSAVEEGTASGSDGNFSEDVLSKFDEWTFIPDSSMDSVFAQSDDLDSEGSEGSFLVKKKSNSISVGEFYRDAVLQRCSPNLQRHSNSLGPIFDHEDLLKRKRKILSSDDSLRSSKLQSHMRHSDSISSLASEREYITSLDLSANELRDIDALSQKCCISVHLEHLEKLELHQNALTSFPQQLCETLKSLTHLDLHSNKFTSFPSYLLKMSCIANLDVSRNDIGPSVVLDPTVKCPTLKQFNLSYNQLSFVPENLTDVVEKLEQLILEGNKISGICSPLRLKELKILNLSKNHISSLSENFLEACPKVESFSARMNFLAAMPFLPPSMTILKLSQNKFSCIPEAILNLPHLRSLDMSSNDIQYLPGPAHWKSLNLRELLFSHNQISILDLSEKAYLWSRVEKLHLSHNKLKEIPPEIGCLENLTSLDVSYNLELRSFPNEMGKLSKIWDLPLDELHLNFDFKHIGCKAKDIIRFLQQRLKKAVPYNRMKLMIVGNTGSGKTTLLQQLMKTKKSDLGMQSATVGIDVKDWPIQIRDKRKRDLVLNVWDFAGREEFYSTHPHFMTQRALYLAVYDLSKGQAEVDAMKPWLFNIKARASSSPVILVGTHLDVSDEKQRKACMSKITKELLNKRGFPAIRDYHFVNATEESDALAKLRKTIINESLNFKIRDQLVVGQLIPDCYVELEKIILSERKNVPIEFPVIDRKRLLQLVRENQLQLDENELPHAVHFLNESGVLLHFQDPALQLSDLYFVEPKWLCKIMAQILTVKVEGCPKHPKGIISRRDVEKFLSKKRKFPKNYMSQYFKLLEKFQIALPIGEEYLLVPSSLSDHRPVIELPHCENSEIIIRLYEMPYFPMGFWSRLINRLLEISPYMLSGRERALRPNRMYWRQGIYLNWSPEAYCLVGSEVLDNHPESFLKITVPSCRKGCILLGQVVDHIDSLMEEWFPGLLEIDICGEGETLLKKWALYSFNDGEEHQKILLDDLMKKAEEGDLLVNPDQPRLTIPISQIAPDLILADLPRNIMLNNDELEFEQAPEFLLGDGSFGSVYRAAYEGEEVAVKIFNKHTSLRLLRQELVVLCHLHHPSLISLLAAGIRPRMLVMELASKGSLDRLLQQDKASLTRTLQHRIALHVADGLRYLHSAMIIYRDLKPHNVLLFTLYPNAAIIAKIADYGIAQYCCRMGIKTSEGTPGFRAPEVARGNVIYNQQADVYSFGLLLYDILTTGGRIVEGLKFPNEFDELEIQGKLPDPVKEYGCAPWPMVEKLIKQCLKENPQERPTSAQVFDILNSAELVCLTRRILLPKNVIVECMVATHHNSRNASIWLGCGHTDRGQLSFLDLNTEGYTSEEVADSRILCLALVHLPVEKESWIVSGTQSGTLLVINTEDGKKRHTLEKMTDSVTCLYCNSFSKQSKQKNFLLVGTADGKLAIFEDKTVKLKGAAPLKILNIGNVSTPLMCLSESTNSTERNVMWGGCGTKIFSFSNDFTIQKLIETRTSQLFSYAAFSDSNIITVVVDTALYIAKQNSPVVEVWDKKTEKLCGLIDCVHFLREVMVKENKESKHKMSYSGRVKTLCLQKNTALWIGTGGGHILLLDLSTRRLIRVIYNFCNSVRVMMTAQLGSLKNVMLVLGYNRKNTEGTQKQKEIQSCLTVWDINLPHEVQNLEKHIEVRKELAEKMRRTSVE imatinib Imatinib small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) Protein kinase inhibitor ponatinib Ponatinib small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) gsk3-xiii GSK3-XIII small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) lrrk2-msn-6 1536356 n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1536358 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false phosphorylation phosphorylation reaction phosphorylation false false false Figure 1,2,3 Table 1. kcat = 8.1/min Ka (ATP) = 69uM Kb = 87uM alpha (Km)= 1.8 The t-I2020T mutant has lower kinase activity than t-WT but ATP binds to the mutant much more tightly than t-WT, with a K_ATP value 40-fold lower than that of t-WT. Incubation at room temperature for 150 min. Initial velocities were measured as a function of [ATP] at a saturating concentration of LRRKtide. lrrk2-msn-7 1536356 n/a 1536358 enzyme enzyme neutral component neutral component Q5S007:p.Ile2020Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false Figure 1,2,3 Table 1.-2 kcat = 4.4/min Ka (ATP) = 1.6uM Kb = 110uM alpha (Km)= 7.3 The t-I2020T mutant has lower kinase activity than t-WT but ATP binds to the mutant much more tightly than t-WT, with a K_ATP value 40-fold lower than that of t-WT. Incubation at room temperature for 150 min. Initial velocities were measured as a function of [ATP] at a saturating concentration of LRRKtide. lrrk2-msn-8 1536356 n/a 1536358 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component P26038:p.Thr558Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false Figure 1, Table 1.-3 kcat = 0.31/min Ka (ATP) = 77uM Kb = 554uM alpha (Km)= 2.6 Unlike LRRKtide, when LRRKtideS is the phosphoryl acceptor, the t-I2020T mutant is more active than t-WT. Incubation at room temperature for 150 min. Initial velocities were measured as a function of [ATP] at a saturating concentration of LRRKtideS. lrrk2-msn-9 1536356 n/a 1536358 enzyme enzyme 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q5S007:p.Ile2020Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P26038:p.Thr558Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false Figure 1,2,3 Table 1.-4 kcat = 0.75min Ka (ATP) = 7.6uM Kb = 263uM alpha (Km)= 1.7 Unlike LRRKtide, when LRRKtideS is the phosphoryl acceptor, the t-I2020T mutant is more active than t-WT. Incubation at room temperature for 150 min. Initial velocities were measured as a function of [ATP] at a saturating concentration of LRRKtideS. imatinib-lrrk2 1536356 n/a 1536359 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1536358 enzyme enzyme neutral component neutral component 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3. Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration. lrrk2-msn-11 1536356 n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1536358 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q5S007:p.Ile2020Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1536360 inhibitor inhibitor neutral component neutral component phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3 Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration. imatinib-lrrk2-1 1536356 n/a 1536358 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1536361 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3. Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration. lrrk2-msn-12 1536356 n/a 1536358 enzyme enzyme 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q5S007:p.Ile2020Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1536359 inhibitor inhibitor neutral component neutral component n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3 Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration. imatinib-lrrk2-2 1536356 n/a 1536358 enzyme enzyme 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 false Q5S007:p.Ile2020Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1536361 inhibitor inhibitor neutral component neutral component n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3 Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration. imatinib-lrrk2-3 1536356 n/a 1536357 enzyme target enzyme target substrate substrate neutral component neutral component region_lrrktide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1536358 enzyme enzyme neutral component neutral component 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1536360 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 1536356 Figure 4 Table 3. Ponatinib and Imatinib show ATP non-competitive inhibition toward t-WT, but inhibit the t-I2020T mutant via an ATP competitive mechanism. GSK3-XIII shows competitive inhibition against both t-WT and the t-I2020T mutant. Initial velocities of LRRK2-catalyzed LRRKtide phosphorylation were measured as a function of inhibitor concentration at varied ATP concentrations and at a fixed LRRKtide concentration.