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/ keller-2011-1 The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. 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 J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation keller-2011-3 The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. 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 J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation keller-2011-4 The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. 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 J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation keller-2011-5 The large extracellular loop of organic cation transporter 1 influences substrate affinity and is pivotal for oligomerization. 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 J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation s22a1_rat Solute carrier family 22 member 1 Slc22a1 Oct1 Organic cation transporter 1 protein protein rat Rattus norvegicus (Rat) Rat MPTVDDVLEQVGEFGWFQKQAFLLLCLISASLAPIYVGIVFLGFTPGHYCQNPGVAELSQRCGWSQAEELNYTVPGLGPSDEASFLSQCMRYEVDWNQSTLDCVDPLSSLVANRSQLPLGPCEHGWVYDTPGSSIVTEFNLVCGDAWKVDLFQSCVNLGFFLGSLVVGYIADRFGRKLCLLVTTLVTSVSGVLTAVAPDYTSMLLFRLLQGMVSKGSWVSGYTLITEFVGSGYRRTTAILYQMAFTVGLVGLAGVAYAIPDWRWLQLAVSLPTFLFLLYYWFVPESPRWLLSQKRTTRAVRIMEQIAQKNGKVPPADLKMLCLEEDASEKRSPSFADLFRTPNLRKHTVILMYLWFSCAVLYQGLIMHVGATGANLYLDFFYSSLVEFPAAFIILVTIDRIGRIYPIAASNLVTGAACLLMIFIPHELHWLNVTLACLGRMGATIVLQMVCLVNAELYPTFIRNLGMMVCSALCDLGGIFTPFMVFRLMEVWQALPLILFGVLGLTAGAMTLLLPETKGVALPETIEEAENLGRRKSKAKENTIYLQVQTGKSSST 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 oct1_oat1 Oct1 Oat1 fusion protein protein protein rat Rattus norvegicus (Rat) Rat MPTVDDVLEQVGEFGWFQKQAFLLLCLISASLAPIYVGIVFLFTAAIPPHHCRPPANANLSKDGGLEAWLPLDKQGQPESCLRFTSPQWGPPFYNGTEANGTRVTEPCIDGWVYDNSTFPSTIVTEWNLVCSHRAFRQLFQSCVNLGFFLGSLVVGYIADRFGRKLCLLVTTLVTSVSGVLTAVAPDYTSMLLFRLLQGMVSKGSWVSGYTLITEFVGSGYRRTTAILYQMAFTVGLVGLAGVAYAIPDWRWLQLAVSLPTFLFLLYYWFVPESPRWLLSQKRTTRAVRIMEQIAQKNGKVPPADLKMLCLEEDASEKRSPSFADLFRTPNLRKHTVILMYLWFSCAVLYQGLIMHVGATGANLYLDFFYSSLVEFPAAFIILVTIDRIGRIYPIAASNLVTGAACLLMIFIPHELHWLNVTLACLGRMGATIVLQMVCLVNAELYPTFIRNLGMMVCSALCDLGGIFTPFMVFRLMEVWQALPLILFGVLGLTAGAMTLLLPETKGVALPETIEEAENLGRRKSKAKENTIYLQVQTGKSSST mpp N-methyl-4-phenylpyridinium small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) slc22a1-slc22a1-3 1241050 n/a 1241054 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 false 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 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1241054 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 false 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 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1241064 1241061 physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 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. 93.3 slc22a1-slc22a6 1241050 n/a 1241054 unspecified role unspecified role bait bait Q63089:p.[Gly159Val;Gly163Val] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 1241055 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 12.3 rOct1-rOct1 93.3% fig 2, table 1 12.3 oct1_oat1-slc22a1 1241050 n/a 1241056 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 false ecoli Escherichia coli (strain K12) n/a 1241054 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 10.7 fig 2, table 1 10.7 oct1_oat1-slc22a6 1241050 n/a 1241055 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 false ecoli Escherichia coli (strain K12) n/a 1241056 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 58.3 fig 2, table 1 58.3 oct1_oat1-oct1_oat-1 1241050 n/a 1241056 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 false ecoli Escherichia coli (strain K12) n/a 1241056 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 61.0 fig 2 61.0 slc22a1-2 1241051 n/a 1241054 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false 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 1241051 n/a 1241054 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false 4 DTT Dimers observed in the presence of CHAP and tetramers in the presence of CHAPS and LMPG extract. slc22a1-slc22a1-9 1241051 n/a 1241055 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1241055 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 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. mpp-oct1-oat1 1241052 n/a 1241056 unspecified role unspecified role neutral component neutral component region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1241057 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 false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false 1241052 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 mpp-slc22a1 1241052 n/a 1241054 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 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1241057 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 false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false 1241052 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-slc22a1-6 1241053 n/a 1241054 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 false 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 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1241054 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 false 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 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) proximity proximity close-contact colocalization neighbourhood interaction false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 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 93.3 oct1_oat1-oct1_oat-1 1241053 n/a 1241056 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 false ecoli Escherichia coli (strain K12) n/a 1241056 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 false ecoli Escherichia coli (strain K12) proximity proximity close-contact colocalization neighbourhood interaction false false false author score author-based confidence author score A score generated by an author, usually only relevant for that particular dataset. 61.0 7 61.0