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/ NKCC2 surface expression in mammalian cells: down-regulation by novel interaction with aldolase B. NKCC2 surface expression in mammalian cells: down-regulation by novel interaction with aldolase B. J. Biol. Chem. (0021-9258) 2007 imex curation Benziane B., Demaretz S., Defontaine N., Zaarour N., Cheval L., Bourgeois S., Klein C., Froissart M., Blanchard A., Paillard M., Gamba G., Houillier P., Laghmani K. Kamel.Laghmani@bhdc.jussieu.fr 23-Feb-2010: Contacted by JYOTI. Human kidney cDNA orchard Only protein-protein interactions mouse-mktal Mouse thick ascending limb cell line mktal Mouse thick ascending limb cell line confocal microscopy confocal microscopy immunostaining immunostaining Benziane B., Demaretz S., Defontaine N., Zaarour N., Cheval L., Bourgeois S., Klein C., Froissart M., Blanchard A., Paillard M., Gamba G., Houillier P., Laghmani K. Kamel.Laghmani@bhdc.jussieu.fr Accepted 2010-MAR-12 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 orchard Only protein-protein interactions imex curation NKCC2 surface expression in mammalian cells: down-regulation by novel interaction with aldolase B. NKCC2 surface expression in mammalian cells: down-regulation by novel interaction with aldolase B. J. Biol. Chem. (0021-9258) 2007 imex curation Benziane B., Demaretz S., Defontaine N., Zaarour N., Cheval L., Bourgeois S., Klein C., Froissart M., Blanchard A., Paillard M., Gamba G., Houillier P., Laghmani K. Kamel.Laghmani@bhdc.jussieu.fr 23-Feb-2010: Contacted by JYOTI. Human kidney cDNA orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Benziane B., Demaretz S., Defontaine N., Zaarour N., Cheval L., Bourgeois S., Klein C., Froissart M., Blanchard A., Paillard M., Gamba G., Houillier P., Laghmani K. Kamel.Laghmani@bhdc.jussieu.fr Accepted 2010-MAR-12 by ORCHARD. 23-Feb-2010: Contacted by JYOTI. Human kidney cDNA J. Biol. Chem. (0021-9258) 2007 orchard Only protein-protein interactions imex curation aldob_human Fructose-bisphosphate aldolase B ALDOB ALDB Liver-type aldolase protein protein human Homo sapiens Human MAHRFPALTQEQKKELSEIAQSIVANGKGILAADESVGTMGNRLQRIKVENTEENRRQFREILFSVDSSINQSIGGVILFHETLYQKDSQGKLFRNILKEKGIVVGIKLDQGGAPLAGTNKETTIQGLDGLSERCAQYKKDGVDFGKWRAVLRIADQCPSSLAIQENANALARYASICQQNGLVPIVEPEVIPDGDHDLEHCQYVTEKVLAAVYKALNDHHVYLEGTLLKPNMVTAGHACTKKYTPEQVAMATVTALHRTVPAAVPGICFLSGGMSEEDATLNLNAINLCPLPKPWKLSFSYGRALQASALAAWGGKAANKEATQEAFMKRAMANCQAAKGQYVHTGSSGAASTQSLFTACYTY DCE314E7AC5586CA s12a1_mouse Solute carrier family 12 member 1 Slc12a1 Nkcc2 Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 2 BSC1 Kidney-specific Na-K-Cl symporter protein protein mouse Mus musculus Mouse MSVSIPSNSVPSSASRFQVHVINEGHGSAAAVGDSADPPHYEETSFGDEAQNRLRISFRPGNQECYDNFLQTGETAKTDTTFHAYDSHTNTYYLQTFGHNTMDAVPKIEYYRNTGSVSGPKVNRPSLLEIHEQLAKNVTVAPGSADRVANGDGMPGDEQAENKEEDMTGVVKFGWVKGVLVRCMLNIWGVMLFIRLSWIVGEAGIGLGVLIILLSTMVTSITGLSTSAIATNGFVRGGGAYYLISRSLGPEFGGSIGLIFAFANAVAVAMYVVGFAETVVDLLKESDSMMVDPTNDIRIIGSITVVILLGISVAGMEWEAKAQVILLVILLIAIANFFIGTVIPSNNEKKSRGFFNYQASIFAENFGPSFTKGEGFFSVFAIFFPAATGILAGANISGDLEDPQDAIPRGTMLAIFITTVAYIGVAICVAACVVRDATGSMNDTIVSGMNCNGSAACGLGYDFSRCQHEPCQYGLMNNFQVMSMVSGFGPLITAGIFSATLSSALASLVSAPKVFQALCKDNIFKGLQFFAKGYGKNNEPLRGYFLTFVIAMAFILIAELNVIAPIISNFFLASYALINFSCFHASYAKSPGWRPAYGIYNMWVSLFGAILCCAVMFVINWWAAVITYVIELFLYIYVTYKKPDVNWGSSTQALSYVSALDNALELTTVEDHVKNFRPQCIVLTGGPMTRPALLDITHAFTKNSGLCICCEVFVGPRKLCVKEMNSGMAKKQAWLIKNKIKAFYAAVAADCFRDGVRSLLQASGLGRMKPNTLVIGYKKNWRKAPLSELENYVGIIHDAFDFEIGVVIVRISQGFDISPVLQVQDELEKLEQERLALEAAIKDNECEEGKGGIRGLFKKAGKLNITKPAPKKDGNISSIQSMHVGEFNQKLVEASAQFKKKQGKGTIDVWWLFDDGGLTLLIPYILTLRKKWKDCKLRIYVGGKITRIEEEKISMASLLSKFRIKFADIHIIGDINIKPNKESWKVFEEMIEPYRLHESHKDLTTAEKLKRESPWKITDAELEAVKEKSYRQVRLNELLQEHSRAANLIVLSLPVARKGSISDLLYMAWLEILTKNLPPVLLVRGNHKNVLTFYS E3E134016ECCE3B0 nkcc2_aldolase 2 3 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 9 physical association physical association Fig. 3C. Yeast two-hybrid analysis identifies an interaction between aldolase B and the C1 peptide of the NKCC2 C-terminal part nkcc2_aldolase-2 1 3 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined didma-okp Opossum kidney cell line okp Opossum kidney cell line Annotated at this level as not clear which opossum was used. Only use as a cell line for transfected proteins, do not annotate endogenous proteins 4 unspecified role unspecified role prey prey region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined didma-okp Opossum kidney cell line okp Opossum kidney cell line Annotated at this level as not clear which opossum was used. Only use as a cell line for transfected proteins, do not annotate endogenous proteins colocalization colocalization Fig. 7 Immunofluorescence confocal microscopy showing distribution of NKCC2 and aldolase B in MKTAL cells.