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 cantarelli-2007-5 yeast Saccharomyces cerevisiae 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 predetermined predetermined participant predetermined Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation cantarelli-2007-1 in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay anti tag western anti tag western blot anti tag western Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation cantarelli-2007-4 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 Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation cantarelli-2007-6 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation cantarelli-2007-7 mouse-fibroblast Mus musculus fibroblast Fibroblast Resident cell of connective tissue, mesodermally derived fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation cantarelli-2007-2 in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Cantarelli VV., Kodama T., Nijstad N., Abolghait SK., Nada S., Okada M., Iida T., Honda T. vcantarelli@weinmann.com.br Cellular microbiology 2007 Only protein-protein interactions imex curation imex curation src8_mouse Src substrate cortactin Cttn Ems1 protein protein mouse Mus musculus Mouse MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDRSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSSAVGFDYKERLAKHESQQDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKKQTPPASPSPQPIEDRPPSSPIYEDAAPFKAEPSYRGSEPEPEYSIEAAGIPEAGSQQGLTYTSEPVYETTEAPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ wasl_rat Neural Wiskott-Aldrich syndrome protein Wasl protein protein rat Rattus norvegicus (Rat) Rat MSSGQQPPRRVTNVGSLLLTPQENESLFSFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDAPNGPNLPMATVDIKNPEITTNRFYSSQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHSSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPGVVVPPPPPNRMYPPPPPALPSSAPSGPPPPPPLSMAGSTAPPPPPPPPPPPGPPPPPGLPSDGDHQVPASSGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVSDGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDDDDEEDFQDDDEWED espfu_eco57 Secreted effector protein EspF(U) EspF-like protein encoded on prophage U Tir-cytoskeleton coupling protein TccP espF(U) tccP Z3072 protein protein eco57 Escherichia coli O157:H7 MINNVSSLFPTVNRNITAVYKKSSFSVSPQKITLNPVKISSPFSPSSSSISATTLFRAPNAHSASFHRQSTAESSLHQQLPNVRQRLIQHLAEHGIXPARSMAEHIPPAPKWPAPPPPVQNEQSRPLPDVAQRLMQHLAEHGIQPARNMAEHIPPAPNWPAPTPPVQNEQSRPLPDVAQRLMQHLAEHGIQPARNMAEHIPPAPXWXAPTPPVQNEQSRPLPDVAQRLMQHLAEHGIZPARSMAEHIPPAPNWPAPPPPVQNEQSRPLPDVAQRLXQHLAEHGIQPARNMAEHIPPAPNWPAPXXPVXNEQSRPLXDVAXRLMQHLAEHGIQPARNMAEHIPPAPNWXAPTPPVQNEQSRPLPDVAQRLMQHLAEHGINTSKRS tir_eco57 Translocated intimin receptor Tir tir espE Secreted effector protein Tir Z5112 ECs4561 protein protein eco57 Escherichia coli O157:H7 MPIGNLGHNPNVNNSIPPAPPLPSQTDGAGGRGQLINSTGPLGSRALFTPVRNSMADSGDNRASDVPGLPVNPMRLAASEITLNDGFEVLHDHGPLDTLNRQIGSSVFRVETQEDGKHIAVGQRNGVETSVVLSDQEYARLQSIDPEGKDKFVFTGGRGGAGHAMVTVASDITEARQRILELLEPKGTGESKGAGESKGVGELRESNSGAENTTETQTSTSTSSLRSDPKLWLALGTVATGLIGLAATGIVQALALTPEPDSPTTTDPDAAASATETATRDQLTKEAFQNPDNQKVNIDELGNAIPSGVLKDDVVANIEEQAKAAGEEAKQQAIENNAQAQKKYDEQQAKRQEELKVSSGAGYGLSGALILGGGIGVAVTAALHRKNQPVEQTTTTTTTTTTTSARTVENKPANNTPAQGNVDTPGSEDTMESRRSSMASTSSTFFDTSSIGTVQNPYADVKTSLHDSQVPTSNSNTSVQNMGNTDSVVYSTIQHPPRDTTDNGARLLGNPSAGIQSTYARLALSGGLRHDMGGLTGGSNSAVNTSNNPPAPGSHRFV src8_human Src substrate cortactin CTTN EMS1 Amplaxin Oncogene EMS1 protein protein human Homo sapiens Human MWKASAGHAVSIAQDDAGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDKSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQRDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSAAVGFDYKEKLAKHESQQDYSKGFGGKYGVQKDRMDKNASTFEDVTQVSSAYQKTVPVEAVTSKTSNIRANFENLAKEKEQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKTQTPPVSPAPQPTEERLPSSPVYEDAASFKAELSYRGPVSGTEPEPVYSMEAADYREASSQQGLAYATEAVYESAEAPGHYPAEDSTYDEYENDLGITAVALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ cttn-wasl-2 3638309 N/A 3638315 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed SH3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Tyr421Asp;Tyr466Asp] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.Trp525Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Ser405Asp;Ser418Asp] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 3638316 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false fig. 4d To further support the results from our Y2H assay, we decided to test the interaction of our cortactin constructs with N‐WASP. N‐WASP specifically interacted with the C‐terminus, but not with the N‐terminus of cortactin (Fig. 4D, grey bars). More importantly, N‐WASP was able to interact with Cort S/D, mimicking serine phosphorylation, but was almost completely unable to interact with the wild type or the construct mimicking tyrosine phosphorylation (Cort Y/D). cttn-espf(u)-3 3638309 N/A 3638315 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed SH3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Tyr421Asp;Tyr466Asp] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Ser405Asp;Ser418Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.Trp525Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false N/A 3638317 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false fig. 4d the Y2H assay confirmed the interaction between EspFu and N‐WASP (top, white bar), as well as between EspFu and the C‐terminus of cortactin (black bars). More importantly, in accordance with the previous results, our Y2H assay showed a strong interaction between EspFu and Cort Y/D form, as compared with the wild‐type or Cort S/D mutant. wasl-espf(u)-3 3638309 N/A 3638316 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed N/A 3638317 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false fig. 4d the Y2H assay confirmed the interaction between EspFu and N‐WASP (top, white bar), as well as between EspFu and the C‐terminus of cortactin (black bars). More importantly, in accordance with the previous results, our Y2H assay showed a strong interaction between EspFu and Cort Y/D form, as compared with the wild‐type or Cort S/D mutant. cttn-1 3638309 N/A 3638318 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3638315 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed SH3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Tyr421Asp;Tyr466Asp] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Ser405Asp;Ser418Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false fig. 5d the Y2H assay confirmed the interaction between EspFu and N‐WASP (top, white bar), as well as between EspFu and the C‐terminus of cortactin (black bars). More importantly, in accordance with the previous results, our Y2H assay showed a strong interaction between EspFu and Cort Y/D form, as compared with the wild‐type or Cort S/D mutant. cttn-espf(u)-1 3638310 N/A 3638317 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 c-term range C-terminal range c-term range c-term range C-terminal range c-term range false myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false eco57 Escherichia coli O157:H7 N/A 3638315 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 sh3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false fig. 4a left espf(u)-cttn-3 3638310 N/A 3638315 unspecified role unspecified role prey prey purified purified ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 N/A 3638318 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 binding region biological feature biological feature certain certain sequence position certain certain certain sequence position certain false eco57 Escherichia coli O157:H7 direct interaction direct interaction false false false fig. 5a espf(u)-cttn-4 3638310 N/A 3638318 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 c-term range C-terminal range c-term range c-term range C-terminal range c-term range false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false eco57 Escherichia coli O157:H7 N/A 3638315 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 sh3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Tyr421Asp;Tyr466Asp] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Ser405Asp;Ser418Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction 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) direct interaction direct interaction false false false fig. 5c we performed a gel overlay assay using several GST‐tagged cortactin constructs immobilized onto PVDF membranes. His‐Tir‐Nt was added as the probe and was found to bind weakly to cortactin wild type or the S/D form (Fig. 5C). Compared with that, the binding of His‐Tir‐Nt to Cort Y/D seemed to be only slightly improved (Fig. 5C, Y/D). Then, we decided to phosphorylate the GST‐Cort wild type in vitro using recombinant c‐Src. As shown in Fig. 5C (wild‐pY), His‐Tir‐Nt reacted strongly with this construct, confirming that tyrosine phosphorylation of cortactin is required to bind the N‐terminus of Tir from EHEC. cttn-espf(u)-2 3638311 N/A 3638319 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3638317 anti tag immunost anti tag immunostaining anti tag immunost unspecified role unspecified role neutral component neutral component infection infection fixed cell fixed cell flag tag 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 colocalization colocalization false false false fig. 4c Infection of HeLa cells with DMEM‐activated EHEC expressing FLAG‐tagged EspFu (as in Fig. 2C and D) resulted in a perfect colocalization of EspFu‐FLAG and tyrosine‐phosphorylated cortactin, which was visible 45 min after infection (Fig. 4C). As the infection proceeded (120 min), cortactin was dephosphorylated and only EspFu‐FLAG was detected near the bacteria. cttn-tir-3 3638311 N/A 3638319 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3638318 unspecified role unspecified role neutral component neutral component infection infection 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 colocalization colocalization false false false fig. 5e Furthermore, using immunofluorescence, Tir and tyrosine‐phosphorylated cortactin were found to colocalize beneath the attached bacteria in HeLa cells infected with DMEM‐activated EHEC for 45 min (Fig. 5E). Accordingly, due to the cortactin dephosphorylation, only Tir was detected in HeLa cells infected with EHEC for 120 min. Infection of HeLa cells with DMEM‐activated EHEC Δtir resulted in a complete failure to induce tyrosine phosphorylation of cortactin, as well as actin assembly (Fig. 5E, Δtir). espf(u)-cttn-2 3638312 N/A 3638317 unspecified role unspecified role bait bait purified purified myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3638315 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false fig. 4b To confirm these findings further, we used lysates obtained from HeLa cells and D5 cells supplemented with purified His‐EspFu‐myc. HeLa cell lysates contain very low levels of tyrosine‐phosphorylated cortactin, in contrast to D5 lysates. As shown in Fig. 4B, tyrosine‐phosphorylated cortactin co‐immunoprecipitates with His‐EspFu‐myc from D5 cells lysates, and no cortactin was co‐immunoprecipitated from the HeLa cells lysates, strongly suggesting that EspFu specifically binds to the tyrosine‐phosphorylated form of cortactin. 1-138 3638312 N/A 3638315 unspecified role unspecified role unspecified role unspecified role cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level phosrestyr421 phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) certain certain sequence position certain certain certain sequence position certain false N/A 3638318 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false eco57 Escherichia coli O157:H7 physical association physical association false false false fig. 5b cttn-espf(u)-4 3638313 N/A 3638317 anti tag immunost anti tag immunostaining anti tag immunost unspecified role unspecified role neutral component neutral component fixed cell fixed cell flag tag 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 N/A 3638315 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization false false false suppl fig 1b when expressed in non‐infected D5 cells, EspFu‐FLAG was found to accumulate in tyrosine‐phosphorylated cortactin‐rich structures (Fig. S1B), again supporting the idea that EspFu associates with tyrosine‐phosphorylated cortactin inside the host cells. cttn-espf(u)-5 3638313 N/A 3638315 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3638317 anti tag immunost anti tag immunostaining anti tag immunost unspecified role unspecified role neutral component neutral component infection infection 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 colocalization colocalization false false false supll fig 2b Infection of HeLa cells with DMEM‐activated EHEC expressing FLAG‐tagged EspFu (as in Fig. 2C and D) resulted in a perfect colocalization of EspFu‐FLAG and tyrosine‐phosphorylated cortactin, which was visible 45 min after infection (Fig. 4C). As the infection proceeded (120 min), cortactin was dephosphorylated and only EspFu‐FLAG was detected near the bacteria. espf(u)-cttn-1 3638314 N/A 3638317 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 c-term range C-terminal range c-term range c-term range C-terminal range c-term range false myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false eco57 Escherichia coli O157:H7 N/A 3638315 unspecified role unspecified role bait bait 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 sh3 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Tyr421Asp;Tyr466Asp] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q60598:p.[Ser405Asp;Ser418Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction 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) direct interaction direct interaction false false false fig. 4a centre