MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Tetrameric structure of SUR2B revealed by electron microscopy of oriented single particles. Tetrameric structure of SUR2B revealed by electron microscopy of oriented single particles. FEBS J. (1742-464X) 2013 Fotinou C., Aittoniemi J., de Wet H., Polidori A., Pucci B., Sansom MS., Vénien-Bryan C., Ashcroft FM. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column experimental particp experimental participant identification experimental particp Fotinou C., Aittoniemi J., de Wet H., Polidori A., Pucci B., Sansom MS., Vénien-Bryan C., Ashcroft FM. FEBS J. (1742-464X) 2013 Tetrameric structure of SUR2B revealed by electron microscopy of oriented single particles. Tetrameric structure of SUR2B revealed by electron microscopy of oriented single particles. FEBS J. (1742-464X) 2013 Fotinou C., Aittoniemi J., de Wet H., Polidori A., Pucci B., Sansom MS., Vénien-Bryan C., Ashcroft FM. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy experimental particp experimental participant identification experimental particp Fotinou C., Aittoniemi J., de Wet H., Polidori A., Pucci B., Sansom MS., Vénien-Bryan C., Ashcroft FM. FEBS J. (1742-464X) 2013 q63563-2 ATP-binding cassette sub-family C member 9 Abcc9 Sulfonylurea receptor 2 Sur2 protein protein rat Rattus norvegicus (Rat) Rat MSLSFCGNNISSYNIYHGVLQNPCFVDALNLVPHVFLLFITFPILFIGWGSQSSKVQIHHNTWLHFPGHNLRWILTFALLFVHVCEIAEGIVSDSQRASRHLHLFMPAVMGFVATTTSIVYYHNIETSNFPKLLLALFLYWVMAFITKTIKLVKYWQLGWGMSDLRFCITGVMVILNGLLMAVEINVIRVRRYVFFMNPQKVKPPEDLQDLGVRFLQPFVNLLSKATYWWMNTLIISAHRKPIDLKAIGKLPIAMRAVTNYVCLKEAYEEQKKKAADHPNRTPSIWLAMYRAFGRPILLSSTFRYLADLLGFAGPLCISGIVQRVNEPKNNTTRFSETLSSKEFLENAHVLAVLLFLALILQRTFLQASYYVTIETGINLRGALLAMIYNKILRLSTSNLSMGEMTLGQINNLVAIETNQLMWFLFLCPNLWAMPVQIIMGVILLYNLLGSSALVGAAVIVLLAPIQYFIATKLAEAQKSTLDYSTERLKKTNEILKGIKLLKLYAWEHIFCKSVEETRMKELSSLKTFALYTSLSIFMNAAIPIAAVLATFVTHAYASGNNLKPAEAFASLSLFHILVTPLFLLSTVVRFAVKAIISVQKLNEFLLSDEIGEDSWRTGEGTLPFESCKKHTGVQSKPINRKQPGRYHLDNYEQARRLRPAETEDVAIKVTNGYFSWGSGLATLSNIDIRIPTGQLTMIVGQVGCGKSSLLLAILGEMQTLEGKVYWNNVNESEPSFEATRSRSRYSVAYAAQKPWLLNATVEENITFGSSFNRQRYKAVTDACSLQPDIDLLPFGDQTEIGERGINLSGGQRQRICVARALYQNTNIVFLDDPFSALDIHLSDHLMQEGILKFLQDDKRTVVLVTHKLQYLTHADWIIAMKDGSVLREGTLKDIQTKDVELYEHWKTLMNRQDQELEKDMEADQTTLERKTLRRAMYSREAKAQMEDEDEEEEEEEDEDDNMSTVMRLRTKMPWKTCWWYLTSGGFFLLFLMIFSKLLKHSVIVAIDYWLATWTSEYSINDPGKADQTFYVAGFSILCGAGIFLCLVTSLTVEWMGLTAAKNLHHNLLNKIILGPIRFFDTTPLGLILNRFSADTNIIDQHIPPTLESLTRSTLLCLSAIGMISYATPVFLIALAPLGVAFYFIQKYFRVASKDLQELDDSTQLPLLCHFSETAEGLTTIRAFRHETRFKQRMLELTDTNNIAYLFLSAANRWLEVRTDYLGACIVLTASIASISGSSNSGLVGLGLLYALTITNYLNWVVRNLADLEVQMGAVKKVNSFLTMESENYEGTMDPSQVPEHWPQEGEIKIHDLCVRYENNLKPVLKHVKAYIKPGQKVGICGRTGSGKSSLSLAFFRMVDIFDGKIVIDGIDISKLPLHTLRSRLSIILQDPILFSGSIRFNLDPECKCTDDRLWEALEIAQLKNMVKSLPGGLDATVTEGGENFSVGQRQLFCLARAFVRKSSILIMDEATASIDMATENILQKVVMTAFADRTVVTIAHRVHTILTADLVIVMKRGNILEYDTPESLLAQEDGVFASFVRADM mint 8666453CED33154D abcc9-1 1 SUR2B 3 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 4.0 direct interaction direct interaction f1b f1c mint Gel filtration revealed that SUR2B eluted as a single peak corresponding to a single oligomeric species (Figure 1B). The calculated molecular weight indicated that this species is likely to be a tetramer. No larger aggregates or other protein species were detected. SDS-PAGE and Western blotting confirmed that the peak corresponds to SUR2B (Figure 1C). On SDS-PAGE gels the protein runs as a protomer. abcc9-1 2 SUR2B 3 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 4.0 direct interaction direct interaction f2 mint Following purification, SUR2B bearing a C-terminal HAT tag was bound to a fluorinated lipid (phenyl-HF-NTA-Ni) monolayer [32], negatively stained and examined by electron microscopy. SUR2B appeared as a homogeneous population of single particles that were square-shaped in cross-section with an unstained central region. Figure 2A shows examples of typical single particles used for analysis. All the particles showed the same orientation, presumably because of unidirectional binding to the lipid monolayer. Figure 2B shows the average of 297 particles, which has a calculated resolution of 21Å. It is evident that this shows clear four-fold rotational symmetry and thus C4 symmetry was subsequently applied (Figure 2C).