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 Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Cellular microbiology 2010 imex curation 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 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-FEB-27 AT 12:56 GMT AT 12:56 GMT by LUANA Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Cellular microbiology 2010 imex curation 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 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-FEB-27 AT 12:56 GMT AT 12:56 GMT by LUANA Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Cellular microbiology 2010 imex curation 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 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-FEB-27 AT 12:56 GMT AT 12:56 GMT by LUANA Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Binding to Na(+) /H(+) exchanger regulatory factor 2 (NHERF2) affects trafficking and function of the enteropathogenic Escherichia coli type III secretion system effectors Map, EspI and NleH. Cellular microbiology 2010 imex curation 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 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-FEB-27 AT 12:56 GMT AT 12:56 GMT by LUANA Cellular microbiology 2010 Only protein-protein interactions imex curation imex curation 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 4F5D341590D22ED7 b7ur60_eco27 E2348C_1442 protein protein eco27 Escherichia coli O127:H6 (strain E2348/69 / EPEC) MNIQPIVTSGITTQNNRHHHAEQTSPTQIPQSELPNGCETGFVVHIPEDMQRHAPECGETTALLSLIKDEGLLSGLDKYLAPHLEEGSAGKKALDMFGLFNVSQMALEIPSTVPGISGKYGVQLNIVKPDIHPTSGNYFLQIFPLHDEIGINFKDLPGPLKNALSNSNIPTTVSTAASTIASATTSTVTTASKDPIPWFGLTAQVVRNHGVELPIVKTENGWKLVGETPLTPDGPKANYTEEWVIRPGEADFKYGASPLQATLGLEFGAHFKWDLDNPNTKYAVLTNAAANALGAVGGFAVSRFTGTDPMLSPHIGAMVGQAAGHAIQYNTPGLKPDTILWWAGTTLGLADLNKAEFGEARFTDYPRIWWHAREGAIFPNKADIEHATGADIRAMEEGVSVGQRHPNPEDVVINIESNNSPHHNPSNYVDTVDIIQETRV 4ABDAE5CBBD4EA55 b7uil3_eco27 Methionine aminopeptidase Peptidase M E2348C_0173 map protein protein eco27 Escherichia coli O127:H6 (strain E2348/69 / EPEC) MAISIKTPEDIEKMRVAGRLAAEVLEMIEPYVKPGVSTGELDRICNDYIVNEQHAVSACLGYHGYPKSVCISINEVVCHGIPDDAKLLKDGDIVNIDVTVIKDGFHGDTSKMFIVGKPTIMGERLCRITQESLYLALRMVKPGINLREIGAAIQKFVEAEGFSVVREYCGHGIGRGFHEEPQVLHYDSRETNVVLKPGMTFTIEPMVNAGKKEIRTMKDGWTVKTKDRSLSAQYEHTIVVTDNGCEILTLRKDDTIPAIISHDE F2D0B57715A67851 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 E33AF87016D37A65 q8x831_eco57 nleH1-1 ECs_0848 E5F08_07760 protein protein eco57 Escherichia coli O157:H7 MLSPYSVNLGCSWNSLTRNLTSPDNRVLSSVRDAAVHSDNGAQVKVGNRTYRVVATDNKFCVTRESHSGCFTNLLHRLGWPKGEISRKIEVMLNASPVSAAMERGIVHSNRPDLPPVDYAPPELPSVDYNRLSVPGNVIGKGGNAVVYEDAEDATKVLKMFTTSQSNEEVTSEVRCFNQYYGAGSAEKIYGNNGDIIGIRMDKINGESLLNISSLPAQAEHAIYDMFDRLEQKGILFVDTTETNVLYDRAKNEFNPIDISSYNVSDRSWSESQIMQSYHGGKQDLISVVLSKI DIP protein Q8X831 original sequence version: 57 slc9a3r1-map-1 1 7 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli 8 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 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 ecolx Escherichia coli direct interaction direct interaction fig. 1a map-slc9a3r2-2 1 7 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli 5 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 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 ecolx Escherichia coli direct interaction direct interaction fig. 1a nleh1_1-slc9a3r2-1 1 9 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli 5 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 ecolx Escherichia coli direct interaction direct interaction fig. 1a map-1 3 7 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 ecolx Escherichia coli 5 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 ecolx Escherichia coli direct interaction direct interaction fig. 1b map-slc9a3r1-1 3 7 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 ecolx Escherichia coli 5 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 pdz2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction fig. 1b b7ur60_eco27-slc9a3r2-1 3 6 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 ecolx Escherichia coli 5 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 ecolx Escherichia coli direct interaction direct interaction fig. 1b b7ur60_eco27-slc9a3r2-2 3 6 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 ecolx Escherichia coli 5 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 pdz2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction fig. 1b nleh1_1-slc9a3r2-2 3 9 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 ecolx Escherichia coli 5 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 ecolx Escherichia coli direct interaction direct interaction fig. 1b b7ur60_eco27-slc9a3r2-3 3 5 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 ecolx Escherichia coli 6 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 ecolx Escherichia coli direct interaction direct interaction fig. 1e slc9a3r2-nleh1_1-1 3 5 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 ecolx Escherichia coli 9 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 ecolx Escherichia coli direct interaction direct interaction fig. 1e slc9a3r2-b7ur60_eco27-1 2 6 unspecified role unspecified role prey prey infection infection 5 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 physical association physical association 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 2 9 unspecified role unspecified role prey prey infection infection 5 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 physical association physical association 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. b7ur60_eco27-slc9a3r2-4 4 6 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 5 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 colocalization colocalization 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 4 9 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 5 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 colocalization colocalization 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 4 7 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 5 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 colocalization colocalization 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).