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 http://mint.bio.uniroma2.it/mint martinez-2010-2 in vivo in vivo far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Martinez E., Schroeder GN., Berger CN., Lee SF., Robinson KS., Badea L., Simpson N., Hall RA., Hartland EL., Crepin VF., Frankel G. g.frankel@imperial.ac.uk Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation martinez-2010-1 in vitro In vitro protein array protein array anti tag western anti tag western blot anti tag western Martinez E., Schroeder GN., Berger CN., Lee SF., Robinson KS., Badea L., Simpson N., Hall RA., Hartland EL., Crepin VF., Frankel G. g.frankel@imperial.ac.uk Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation martinez-2010-4 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Martinez E., Schroeder GN., Berger CN., Lee SF., Robinson KS., Badea L., Simpson N., Hall RA., Hartland EL., Crepin VF., Frankel G. g.frankel@imperial.ac.uk Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation martinez-2010-3 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Martinez E., Schroeder GN., Berger CN., Lee SF., Robinson KS., Badea L., Simpson N., Hall RA., Hartland EL., Crepin VF., Frankel G. g.frankel@imperial.ac.uk Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation b7uil3_eco27 Methionine aminopeptidase Peptidase M E2348C_0173 map protein protein eco27 Escherichia coli O127:H6 (strain E2348/69 / EPEC) MAISIKTPEDIEKMRVAGRLAAEVLEMIEPYVKPGVSTGELDRICNDYIVNEQHAVSACLGYHGYPKSVCISINEVVCHGIPDDAKLLKDGDIVNIDVTVIKDGFHGDTSKMFIVGKPTIMGERLCRITQESLYLALRMVKPGINLREIGAAIQKFVEAEGFSVVREYCGHGIGRGFHEEPQVLHYDSRETNVVLKPGMTFTIEPMVNAGKKEIRTMKDGWTVKTKDRSLSAQYEHTIVVTDNGCEILTLRKDDTIPAIISHDE nhrf2_human Na(+)/H(+) exchange regulatory cofactor NHE-RF2 SLC9A3R2 NHERF2 Tyrosine kinase activator protein 1 SRY-interacting protein 1 Solute carrier family 9 isoform A3 regulatory factor 2 NHE3 kinase A regulatory protein E3KARP Sodium-hydrogen exchanger regulatory factor 2 protein protein human Homo sapiens Human MAAPEPLRPRLCRLVRGEQGYGFHLHGEKGRRGQFIRRVEPGSPAEAAALRAGDRLVEVNGVNVEGETHHQVVQRIKAVEGQTRLLVVDQETDEELRRRQLTCTEEMAQRGLPPAHDPWEPKPDWAHTGSHSSEAGKKDVSGPLRELRPRLCHLRKGPQGYGFNLHSDKSRPGQYIRSVDPGSPAARSGLRAQDRLIEVNGQNVEGLRHAEVVASIKAREDEARLLVVDPETDEHFKRLRVTPTEEHVEGPLPSPVTNGTSPAQLNGGSACSSRSDLPGSDKDTEDGSAWKQDPFQESGLHLSPTAAEAKEKARAMRVNKRAPQMDWNRKREIFSNF b7ur60_eco27 E2348C_1442 protein protein eco27 Escherichia coli O127:H6 (strain E2348/69 / EPEC) MNIQPIVTSGITTQNNRHHHAEQTSPTQIPQSELPNGCETGFVVHIPEDMQRHAPECGETTALLSLIKDEGLLSGLDKYLAPHLEEGSAGKKALDMFGLFNVSQMALEIPSTVPGISGKYGVQLNIVKPDIHPTSGNYFLQIFPLHDEIGINFKDLPGPLKNALSNSNIPTTVSTAASTIASATTSTVTTASKDPIPWFGLTAQVVRNHGVELPIVKTENGWKLVGETPLTPDGPKANYTEEWVIRPGEADFKYGASPLQATLGLEFGAHFKWDLDNPNTKYAVLTNAAANALGAVGGFAVSRFTGTDPMLSPHIGAMVGQAAGHAIQYNTPGLKPDTILWWAGTTLGLADLNKAEFGEARFTDYPRIWWHAREGAIFPNKADIEHATGADIRAMEEGVSVGQRHPNPEDVVINIESNNSPHHNPSNYVDTVDIIQETRV q8x831_eco57 nleH1-1 ECs_0848 E5F08_07760 protein protein eco57 Escherichia coli O157:H7 MLSPYSVNLGCSWNSLTRNLTSPDNRVLSSVRDAAVHSDNGAQVKVGNRTYRVVATDNKFCVTRESHSGCFTNLLHRLGWPKGEISRKIEVMLNASPVSAAMERGIVHSNRPDLPPVDYAPPELPSVDYNRLSVPGNVIGKGGNAVVYEDAEDATKVLKMFTTSQSNEEVTSEVRCFNQYYGAGSAEKIYGNNGDIIGIRMDKINGESLLNISSLPAQAEHAIYDMFDRLEQKGILFVDTTETNVLYDRAKNEFNPIDISSYNVSDRSWSESQIMQSYHGGKQDLISVVLSKI nhrf1_human Na(+)/H(+) exchange regulatory cofactor NHE-RF1 SLC9A3R1 NHERF NHERF1 Ezrin-radixin-moesin-binding phosphoprotein 50 Regulatory cofactor of Na(+)/H(+) exchanger Sodium-hydrogen exchanger regulatory factor 1 Solute carrier family 9 isoform A3 regulatory factor 1 protein protein human Homo sapiens Human MSADAAAGAPLPRLCCLEKGPNGYGFHLHGEKGKLGQYIRLVEPGSPAEKAGLLAGDRLVEVNGENVEKETHQQVVSRIRAALNAVRLLVVDPETDEQLQKLGVQVREELLRAQEAPGQAEPPAAAEVQGAGNENEPREADKSHPEQRELRPRLCTMKKGPSGYGFNLHSDKSKPGQFIRSVDPDSPAEASGLRAQDRIVEVNGVCMEGKQHGDVVSAIRAGGDETKLLVVDRETDEFFKKCRVIPSQEHLNGPLPVPFTNGEIQKENSREALAEAALESPRPALVRSASSDTSEELNSQDSPPKQDSTAPSSTSSSDPILDFNISLAMAKERAHQKRSSKRAPQMDWSKKNELFSNL map-1 3671074 N/A 3671078 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role prey prey purified purified his tag 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 ecolx Escherichia coli direct interaction direct interaction false false false fig. 1b map-slc9a3r1-1 3671074 N/A 3671078 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role prey prey purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false pdz2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1b b7ur60_eco27-slc9a3r2-1 3671074 N/A 3671080 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role prey prey purified purified his tag 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 ecolx Escherichia coli direct interaction direct interaction false false false fig. 1b b7ur60_eco27-slc9a3r2-2 3671074 N/A 3671080 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role prey prey purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false pdz2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1b nleh1_1-slc9a3r2-2 3671074 N/A 3671081 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role prey prey purified purified his tag 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 ecolx Escherichia coli direct interaction direct interaction false false false fig. 1b b7ur60_eco27-slc9a3r2-3 3671074 N/A 3671079 unspecified role unspecified role bait bait purified purified his tag 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 ecolx Escherichia coli N/A 3671080 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified his tag 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 ecolx Escherichia coli direct interaction direct interaction false false false fig. 1e slc9a3r2-nleh1_1-1 3671074 N/A 3671079 unspecified role unspecified role bait bait purified purified his tag 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 ecolx Escherichia coli N/A 3671081 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified gst tag 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 false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1e slc9a3r1-map-1 3671075 N/A 3671078 unspecified role unspecified role prey prey purified purified GST tag 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 false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli N/A 3671082 unspecified role unspecified role bait bait purified purified PDZ1 sufficient to bind sufficient binding region sufficient to bind inferred by curator inferred by curator modelled by curator modeled by curator certain certain sequence position certain certain certain sequence position certain false PDZ2 sufficient to bind sufficient binding region sufficient to bind inferred by curator inferred by curator modelled by curator modeled by curator certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1a map-slc9a3r2-2 3671075 N/A 3671078 unspecified role unspecified role prey prey purified purified GST tag 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 false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role bait bait purified purified PDZ1 sufficient to bind sufficient binding region sufficient to bind inferred by curator inferred by curator modelled by curator modeled by curator certain certain sequence position certain certain certain sequence position certain false PDZ2 sufficient to bind sufficient binding region sufficient to bind inferred by curator inferred by curator modelled by curator modeled by curator certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1a nleh1_1-slc9a3r2-1 3671075 N/A 3671081 unspecified role unspecified role prey prey purified purified GST tag 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 false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli N/A 3671079 unspecified role unspecified role bait bait purified purified PDZ2 sufficient to bind sufficient binding region sufficient to bind inferred by curator inferred by curator modelled by curator modeled by curator certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli direct interaction direct interaction false false false fig. 1a b7ur60_eco27-slc9a3r2-4 3671076 N/A 3671080 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3671079 unspecified role unspecified role neutral component neutral component engineered engineered fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false fig. 3 suppl fig.2 In order to determine if the effectors colocalize with NHERF2, HeLa–NHERF2 cells were transfected with pRK5 (Fig. 3) or pRK5 expressing myc-tagged Map, MapΔC3, NleH1 and NleH1ΔC4 as well as pGFP-EspI and pGFP-EspIΔC7. Analysing the transfected cells by immunofluorescence revealed that Map colocalized with mitochondria (Fig. S1) and NHERF2 (Fig. 3). Plasma membrane-associated EspI and NleH1 colocalized with NHERF2 (Fig. 3). We observed no colocalization of NHERF2 and MapΔC3, EspIΔC7 or NleH1ΔC4 (Fig. S2). nleh1_1-slc9a3r2-3 3671076 N/A 3671081 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3671079 unspecified role unspecified role neutral component neutral component engineered engineered fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false fig. 3; fig. 6a In order to determine if the effectors colocalize with NHERF2, HeLa–NHERF2 cells were transfected with pRK5 (Fig. 3) or pRK5 expressing myc-tagged Map, MapΔC3, NleH1 and NleH1ΔC4 as well as pGFP-EspI and pGFP-EspIΔC7. Analysing the transfected cells by immunofluorescence revealed that Map colocalized with mitochondria (Fig. S1) and NHERF2 (Fig. 3). Plasma membrane-associated EspI and NleH1 colocalized with NHERF2 (Fig. 3). We observed no colocalization of NHERF2 and MapΔC3, EspIΔC7 or NleH1ΔC4 (Fig. S2). map-slc9a3r2-3 3671076 N/A 3671078 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3671079 unspecified role unspecified role neutral component neutral component engineered engineered fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false fig. 3 suppl fig. 1 In order to determine if the effectors colocalize with NHERF2, HeLa–NHERF2 cells were transfected with pRK5 (Fig. 3) or pRK5 expressing myc-tagged Map, MapΔC3, NleH1 and NleH1ΔC4 as well as pGFP-EspI and pGFP-EspIΔC7. Analysing the transfected cells by immunofluorescence revealed that Map colocalized with mitochondria (Fig. S1) and NHERF2 (Fig. 3). Plasma membrane-associated EspI and NleH1 colocalized with NHERF2 (Fig. 3). We observed no colocalization of NHERF2 and MapΔC3, EspIΔC7 or NleH1ΔC4 (Fig. S2). slc9a3r2-b7ur60_eco27-1 3671077 N/A 3671080 unspecified role unspecified role prey prey infection infection N/A 3671079 unspecified role unspecified role bait bait engineered engineered ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false fig.2a Probing infected cell extracts with anti-EspI and anti NleH1 revealed that while both full-length EspI and EspIΔC7 were found in equal amounts, NleH1ΔC4 was unstable (Fig. 2A and B). Following cell lysis the supernatants were subjected to co-immunoprecipitation with anti-HA antibodies. While NHERF2 did not co-immunoprecipitate EspIΔC7 (Fig. 2A) and NleH1ΔC4 (Fig. 2B), both full-length EspI (Fig. 2A) and NleH1 (Fig. 2B) were co-immunoprecipitated. slc9a3r2-nleh1_1-2 3671077 N/A 3671081 unspecified role unspecified role prey prey infection infection N/A 3671079 unspecified role unspecified role bait bait engineered engineered ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false fig.2b Probing infected cell extracts with anti-EspI and anti NleH1 revealed that while both full-length EspI and EspIΔC7 were found in equal amounts, NleH1ΔC4 was unstable (Fig. 2A and B). Following cell lysis the supernatants were subjected to co-immunoprecipitation with anti-HA antibodies. While NHERF2 did not co-immunoprecipitate EspIΔC7 (Fig. 2A) and NleH1ΔC4 (Fig. 2B), both full-length EspI (Fig. 2A) and NleH1 (Fig. 2B) were co-immunoprecipitated.