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/ G2385R and I2020T Mutations Increase LRRK2 GTPase Activity. G2385R and I2020T Mutations Increase LRRK2 GTPase Activity. BioMed research international 2015 imex curation Ho DH., Jang J., Joe EH., Son I., Seo H., Seol W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. wseolha@gmail.com,hseo@hanyang.ac.kr 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Ho DH., Jang J., Joe EH., Son I., Seo H., Seol W. Parkinsons - Interactions investigated in the context of Parkinsons disease LRRK2 proteins obtained from Creative Biomart (I2020T and G2385R) were N-terminally GST tagged and expressed in Sf9 cells. The WT and other mutants were obtained from Invitrogen/Thermoscientific and the expression system was "Baculovirus" and no tag location is provided. sorry- yet more LRRK2 autophos-es! A GTPAse assay kit from Innova Bioscience was used; phosphate ions liberated during the reaction are measured colourimetrically after formation of colored complexes between inorganic phosphate ions and molecules of an unknown dye. wseolha@gmail.com,hseo@hanyang.ac.kr Accepted 2016-AUG-25 AT 10:46 BST AT 10:46 BST by ORCHARD BioMed research international 2015 Only protein-protein interactions imex curation imex curation G2385R and I2020T Mutations Increase LRRK2 GTPase Activity. G2385R and I2020T Mutations Increase LRRK2 GTPase Activity. BioMed research international 2015 imex curation Ho DH., Jang J., Joe EH., Son I., Seo H., Seol W. Parkinsons - Interactions investigated in the context of Parkinsons disease Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. wseolha@gmail.com,hseo@hanyang.ac.kr 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase predetermined predetermined participant predetermined Ho DH., Jang J., Joe EH., Son I., Seo H., Seol W. Parkinsons - Interactions investigated in the context of Parkinsons disease LRRK2 proteins obtained from Creative Biomart (I2020T and G2385R) were N-terminally GST tagged and expressed in Sf9 cells. The WT and other mutants were obtained from Invitrogen/Thermoscientific and the expression system was "Baculovirus" and no tag location is provided. A GTPAse assay kit from Innova Bioscience was used; phosphate ions liberated during the reaction are measured colourimetrically after formation of colored complexes between inorganic phosphate ions and molecules of an unknown dye. wseolha@gmail.com,hseo@hanyang.ac.kr Accepted 2016-AUG-25 AT 10:46 BST AT 10:46 BST by ORCHARD BioMed research international 2015 Only protein-protein interactions imex curation imex curation 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 rab5b_human Ras-related protein Rab-5B RAB5B protein protein human Homo sapiens Human MTSRSTARPNGQPQASKICQFKLVLLGESAVGKSSLVLRFVKGQFHEYQESTIGAAFLTQSVCLDDTTVKFEIWDTAGQERYHSLAPMYYRGAQAAIVVYDITNQETFARAKTWVKELQRQASPSIVIALAGNKADLANKRMVEYEEAQAYADDNSLLFMETSAKTAMNVNDLFLAIAKKLPKSEPQNLGGAAGRSRGVDLHEQSQQNKSQCCSN D47AD834BCF8B591 gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 gtp-lrrk2-8 2 5 enzyme target enzyme target substrate substrate neutral component neutral component 3 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Asp1994Ala mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 16:12:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1994ala' Tue Jun 30 17:59:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1994Ala' Q5S007:p.Gly2019Ser mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain G S Wed May 02 19:02:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 17:58:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' Q5S007:p.Arg1441Cys mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R C Wed May 02 17:10:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441cys' Tue Jun 30 18:01:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Cys' gtpase reaction gtpase reaction 1 Proteins obtained from Thermoscientific (Invitrogen) gtp-lrrk2-9 2 5 enzyme target enzyme target substrate substrate neutral component neutral component 3 enzyme enzyme neutral component neutral component n-terminal 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 Q5S007:p.Ile2020Thr mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain I T Wed May 02 14:40:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile2020thr' Tue Jun 30 16:58:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile2020Thr' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Gly2385Arg mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain G R Wed May 02 15:03:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2385arg' Tue Jun 30 18:19:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2385Arg' 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 gtpase reaction gtpase reaction 1-2 Proteins obtained from Creative Biomart (Shirley, NY, USA). gtp-lrrk2-15 1 4 enzyme target enzyme target substrate substrate neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 3 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Asp1994Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 16:07:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1994ala' Tue Jun 30 15:38:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1994Ala' 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 18:14:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 18:11:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' Q5S007:p.Arg1441Cys mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R C Wed May 02 15:21:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441cys' Tue Jun 30 16:01:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Cys' phosphorylation phosphorylation reaction phosphorylation 2 Proteins obtained from Thermoscientific (Invitrogen) gtp-lrrk2-16 1 4 enzyme target enzyme target substrate substrate neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 3 enzyme enzyme neutral component neutral component n-terminal 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 Q5S007:p.Ile2020Thr mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain I T Wed May 02 16:35:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile2020thr' Tue Jun 30 16:56:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile2020Thr' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Gly2385Arg mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain G R Wed May 02 18:26:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2385arg' Tue Jun 30 18:22:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2385Arg' 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 2 Proteins obtained from Creative Biomart (Shirley, NY, USA). lrrk2-rab5b-10 1 3 putative self putative self putative self putative self 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Asp1994Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:36:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1994ala' Tue Jun 30 20:17:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1994Ala' 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 15:59:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2019ser' Tue Jun 30 18:30:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2019Ser' Q5S007:p.Arg1441Cys mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R C Wed May 02 18:01:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1441cys' Tue Jun 30 16:00:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1441Cys' phosphorylation phosphorylation reaction phosphorylation true 2a Proteins obtained from Thermoscientific (Invitrogen) lrrk2-rab5b-11 1 3 putative self putative self putative self putative self n-terminal 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 Q5S007:p.Ile2020Thr mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain I T Wed May 02 16:08:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile2020thr' Tue Jun 30 16:12:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile2020Thr' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q5S007:p.Gly2385Arg mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain G R Wed May 02 19:13:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly2385arg' Tue Jun 30 16:52:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly2385Arg' 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 2b Proteins obtained from Creative Biomart (Shirley, NY, USA).