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/ The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. J. Biol. Chem. (0021-9258) 2011 imex curation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de Accepted 2011-NOV-30 by MGT 14-Mar-2012: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. J. Biol. Chem. (0021-9258) 2011 imex curation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. The authors have calculated the coprecipitation of the FLAG tagged construct as a percentage of the total. This gives an idea of the strength of interaction. Superdex 200 used to determine complex size. Hermann@Koepsell.de Accepted 2011-NOV-30 by MGT J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. J. Biol. Chem. (0021-9258) 2011 imex curation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag visualisation tag visualisation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de Accepted 2011-NOV-30 by MGT 14-Mar-2012: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. J. Biol. Chem. (0021-9258) 2011 imex curation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column tag visualisation tag visualisation Keller T., Egenberger B., Gorboulev V., Bernhard F., Uzelac Z., Gorbunov D., Wirth C., Koppatz S., Dotsch V., Hunte C., Sitte H.H., Koepsell H. Superdex 200 used to determine complex size. Hermann@Koepsell.de Accepted 2011-NOV-30 by MGT 14-Mar-2012: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation s22a6_rat Solute carrier family 22 member 6 Slc22a6 Oat1 Organic anion transporter 1 renal organic anion transporter 1 protein protein rat Rattus norvegicus (Rat) Rat MAFNDLLKQVGGVGRFQLIQVTMVVAPLLLMASHNTLQNFTAAIPPHHCRPPANANLSKDGGLEAWLPLDKQGQPESCLRFTSPQWGPPFYNGTEANGTRVTEPCIDGWVYDNSTFPSTIVTEWNLVCSHRAFRQLAQSLYMVGVLLGAMVFGYLADRLGRRKVLILNYLQTAVSGTCAAYAPNYTVYCVFRLLSGMSLASIAINCMTLNVEWMPIHTRAYVGTLIGYVYSLGQFLLAGIAYAVPHWRHLQLVVSVPFFIAFIYSWFFIESARWYSSSGRLDLTLRALQRVARINGKQEEGAKLSIEVLRTSLQKELTLSKGQASAMELLRCPTLRHLFLCLSMLWFATSFAYYGLVMDLQGFGVSMYLIQVIFGAVDLPAKFVCFLVINSMGRRPAQMASLLLAGICILVNGIIPKSHTIIRTSLAVLGKGCLASSFNCIFLYTGELYPTVIRQTGLGMGSTMARVGSIVSPLVSMTAEFYPSMPLFIFGAVPVVASAVTALLPETLGQPLPDTVQDLKSRSRGKQNQQQQEQQKQMMPLQASTQEKNGL 8BA47BE628324BF2 oct1_oat1 Oct1 Oat1 fusion protein protein protein rat Rattus norvegicus (Rat) Rat MPTVDDVLEQVGEFGWFQKQAFLLLCLISASLAPIYVGIVFLFTAAIPPHHCRPPANANLSKDGGLEAWLPLDKQGQPESCLRFTSPQWGPPFYNGTEANGTRVTEPCIDGWVYDNSTFPSTIVTEWNLVCSHRAFRQLFQSCVNLGFFLGSLVVGYIADRFGRKLCLLVTTLVTSVSGVLTAVAPDYTSMLLFRLLQGMVSKGSWVSGYTLITEFVGSGYRRTTAILYQMAFTVGLVGLAGVAYAIPDWRWLQLAVSLPTFLFLLYYWFVPESPRWLLSQKRTTRAVRIMEQIAQKNGKVPPADLKMLCLEEDASEKRSPSFADLFRTPNLRKHTVILMYLWFSCAVLYQGLIMHVGATGANLYLDFFYSSLVEFPAAFIILVTIDRIGRIYPIAASNLVTGAACLLMIFIPHELHWLNVTLACLGRMGATIVLQMVCLVNAELYPTFIRNLGMMVCSALCDLGGIFTPFMVFRLMEVWQALPLILFGVLGLTAGAMTLLLPETKGVALPETIEEAENLGRRKSKAKENTIYLQVQTGKSSST s22a1_rat Solute carrier family 22 member 1 Slc22a1 Oct1 Organic cation transporter 1 protein protein rat Rattus norvegicus (Rat) Rat MPTVDDVLEQVGEFGWFQKQAFLLLCLISASLAPIYVGIVFLGFTPGHYCQNPGVAELSQRCGWSQAEELNYTVPGLGPSDEASFLSQCMRYEVDWNQSTLDCVDPLSSLVANRSQLPLGPCEHGWVYDTPGSSIVTEFNLVCGDAWKVDLFQSCVNLGFFLGSLVVGYIADRFGRKLCLLVTTLVTSVSGVLTAVAPDYTSMLLFRLLQGMVSKGSWVSGYTLITEFVGSGYRRTTAILYQMAFTVGLVGLAGVAYAIPDWRWLQLAVSLPTFLFLLYYWFVPESPRWLLSQKRTTRAVRIMEQIAQKNGKVPPADLKMLCLEEDASEKRSPSFADLFRTPNLRKHTVILMYLWFSCAVLYQGLIMHVGATGANLYLDFFYSSLVEFPAAFIILVTIDRIGRIYPIAASNLVTGAACLLMIFIPHELHWLNVTLACLGRMGATIVLQMVCLVNAELYPTFIRNLGMMVCSALCDLGGIFTPFMVFRLMEVWQALPLILFGVLGLTAGAMTLLLPETKGVALPETIEEAENLGRRKSKAKENTIYLQVQTGKSSST 9F42131CCCEC0920 mpp N-methyl-4-phenylpyridinium small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) slc22a1-slc22a1-3 2 7 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q63089:p.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Fri Jul 22 11:02:51 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'cys143ser,cys122ser,cys103ser,cys89ser,cys62ser,cys50ser' Wed May 02 17:31:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys50ser,cys62ser,cys89ser,cys103ser,cys122ser,cys143ser' Tue Jun 30 19:31:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser]' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait 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 Q63089:p.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 16:13:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys50ser,cys62ser,cys89ser,cys103ser,cys122ser,cys143ser' 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.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser]' ecoli Escherichia coli (strain K12) 12 15 physical association physical association author score author-based confidence author score 93.3 fig 1, fig 3a, table 1 21.7 % of FLAG protein binding of the 6 cys to ser mutant construct observed. 86.3 % of FLAG protein binding of L447Y/Q448E mutant construct observed. slc22a1-slc22a6 2 7 unspecified role unspecified role bait bait Q63089:p.[Gly159Val;Gly163Val] mutation mutation certain certain sequence position certain certain certain sequence position certain G V certain certain sequence position certain certain certain sequence position certain G V Fri Jul 22 10:57:16 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'gly163val,gly159val' Wed May 02 18:44:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly159val,gly163val' Tue Jun 30 16:48:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly159Val;Gly163Val]' 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association author score author-based confidence author score 12.3 rOct1-rOct1 93.3% fig 2, table 1 oct1_oat1-slc22a1 2 6 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association author score author-based confidence author score 10.7 fig 2, table 1 oct1_oat1-slc22a6 2 5 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association author score author-based confidence author score 58.3 fig 2, table 1 oct1_oat1-oct1_oat-1 2 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association author score author-based confidence author score 61.0 fig 2 slc22a1-2 1 7 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction 4 DTT Dimers observed in the presence of CHAP and tetramers int he presence of CHAPS and LMPG extract. Monomers of Oct1 comigrated with 75kda while dimers comigrated at 158 KDa. slc22a1-1 1 7 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction 4 DTT Dimers observed in the presence of CHAP and tetramers in the presence of CHAPS and LMPG extract. mpp-oct1-oat1 4 6 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figs. 5, 6 table 2 rOct1(Oat1-l) Turnover=8.2, Km =1.1E-4, Inhibition IC50 in presence of TEA Km=1.7E-4, TBuA Km=2.5E-5, TPeA Km=3.6E-6 slc22a1-slc22a1-6 3 7 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q63089:p.[Cys50Ser;Arg61Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain R S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:28:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys50ser,arg61ser,cys89ser,cys103ser,cys122ser,cys143ser' Tue Jun 30 16:14:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys50Ser;Arg61Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser]' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q63089:p.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Fri Jul 22 11:14:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'cys143ser,cys122ser,cys103ser,cys89ser,cys62ser,cys50ser' Wed May 02 19:36:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys50ser,cys62ser,cys89ser,cys103ser,cys122ser,cys143ser' Tue Jun 30 18:56:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys50Ser;Cys62Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser]' ecoli Escherichia coli (strain K12) proximity proximity close-contact colocalization neighbourhood interaction author score author-based confidence author score 93.3 7,8 21.7 % of FLAG protein binding of the 6 cys to ser mutant construct observed. 86.3 % of FLAG protein binding of L447Y/Q448E mutant construct observed. DTT oct1_oat1-oct1_oat-1 3 6 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) proximity proximity close-contact colocalization neighbourhood interaction author score author-based confidence author score 61.0 7 mpp-slc22a1 4 7 unspecified role unspecified role neutral component neutral component Q63089:p.[Cys50Ser;Arg61Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Fri Jul 22 11:14:15 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg61ser,cys89ser,cys103ser,cys122ser,cys143ser,cys50ser' Wed May 02 17:03:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys50ser,arg61ser,cys89ser,cys103ser,cys122ser,cys143ser' Tue Jun 30 18:21:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys50Ser;Arg61Ser;Cys89Ser;Cys103Ser;Cys122Ser;Cys143Ser]' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figs. 5, 6 table 2 rOct1 Turnover=19.5, Km =3.5E-5, Inhibition IC50 in presence of TEA Km=1.5E-4, TBuA Km=1.8E-5, TPeA Km=2.8E-6. roct 6cystoser mutation Turnover=9.1, Km =1.0E-4, Inhibition IC50 in presence of TEA Km=1.8E-4, TBuA Km=2.7E-5, TPeA Km=3.7E-6 slc22a1-slc22a1-9 1 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Nmers eluting in the void volumeobserved in the presence of CHAPS and LMPG extract. DTT supplemental fig. 1 The complex eluted in the void volume indicating a n-nary complex that was not resolved as dimer or tetramer on the sephadex 200 column.