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/ http://www.ebi.ac.uk/ fraipont-2011-3 The integral membrane FtsW protein and peptidoglycan synthase PBP3 form a subcomplex in Escherichia coli. ecoli Escherichia coli (strain K12) adenylate cyclase adenylate cyclase complementation bacterial two-hybrid bacterial two-hybrid adenylate cyclase predetermined predetermined participant predetermined Fraipont C., Alexeeva S., Wolf B., van der Ploeg R., Schloesser M., den Blaauwen T., Nguyen-Disteche M. mng.disteche@ulg.ac.be, t.denblaauwen@uva.nl Microbiology (Reading, Engl.) (1350-0872) 2011 Only protein-protein interactions imex curation PBP3 is also called FtsI fraipont-2011-1 The integral membrane FtsW protein and peptidoglycan synthase PBP3 form a subcomplex in Escherichia coli. ecoli Escherichia coli (strain K12) fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Fraipont C., Alexeeva S., Wolf B., van der Ploeg R., Schloesser M., den Blaauwen T., Nguyen-Disteche M. mng.disteche@ulg.ac.be, t.denblaauwen@uva.nl Microbiology (Reading, Engl.) (1350-0872) 2011 Only protein-protein interactions imex curation PBP3 is also called FtsI fraipont-2011-2 The integral membrane FtsW protein and peptidoglycan synthase PBP3 form a subcomplex in Escherichia coli. ecoli Escherichia coli (strain K12) anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Fraipont C., Alexeeva S., Wolf B., van der Ploeg R., Schloesser M., den Blaauwen T., Nguyen-Disteche M. mng.disteche@ulg.ac.be, t.denblaauwen@uva.nl Microbiology (Reading, Engl.) (1350-0872) 2011 Only protein-protein interactions imex curation PBP3 is also called FtsI ftsi_ecoli Peptidoglycan D,D-transpeptidase FtsI ftsI pbpB b0084 Penicillin-binding protein 3 JW0082 Essential cell division protein FtsI Murein transpeptidase Peptidoglycan synthase FtsI protein protein ecoli Escherichia coli (strain K12) MKAAAKTQKPKRQEEHANFISWRFALLCGCILLALAFLLGRVAWLQVISPDMLVKEGDMRSLRVQQVSTSRGMITDRSGRPLAVSVPVKAIWADPKEVHDAGGISVGDRWKALANALNIPLDQLSARINANPKGRFIYLARQVNPDMADYIKKLKLPGIHLREESRRYYPSGEVTAHLIGFTNVDSQGIEGVEKSFDKWLTGQPGERIVRKDRYGRVIEDISSTDSQAAHNLALSIDERLQALVYRELNNAVAFNKAESGSAVLVDVNTGEVLAMANSPSYNPNNLSGTPKEAMRNRTITDVFEPGSTVKPMVVMTALQRGVVRENSVLNTIPYRINGHEIKDVARYSELTLTGVLQKSSNVGVSKLALAMPSSALVDTYSRFGLGKATNLGLVGERSGLYPQKQRWSDIERATFSFGYGLMVTPLQLARVYATIGSYGIYRPLSITKVDPPVPGERVFPESIVRTVVHMMESVALPGGGGVKAAIKGYRIAIKTGTAKKVGPDGRYINKYIAYTAGVAPASQPRFALVVVINDPQAGKYYGGAVSAPVFGAIMGGVLRTMNIEPDALTTGDKNEFVINQGEGTGGRS This sequence has been withdrawn from Uniprot. ftsw_ecoli Probable peptidoglycan glycosyltransferase FtsW ftsW Cell division protein FtsW Cell wall polymerase Lipid II flippase FtsW Peptidoglycan polymerase JW0087 b0089 protein protein ecoli Escherichia coli (strain K12) MRLSLPRLKMPRLPGFSILVWISTALKGWVMGSREKDTDSLIMYDRTLLWLTFGLAAIGFIMVTSASMPIGQRLTNDPFFFAKRDGVYLILAFILAIITLRLPMEFWQRYSATMLLGSIILLMIVLVVGSSVKGASRWIDLGLLRIQPAELTKLSLFCYIANYLVRKGDEVRNNLRGFLKPMGVILVLAVLLLAQPDLGTVVVLFVTTLAMLFLAGAKLWQFIAIIGMGISAVVLLILAEPYRIRRVTAFWNPWEDPFGSGYQLTQSLMAFGRGELWGQGLGNSVQKLEYLPEAHTDFIFAIIGEELGYVGVVLALLMVFFVAFRAMSIGRKALEIDHRFSGFLACSIGIWFSFQALVNVGAAAGMLPTKGLTLPLISYGGSSLLIMSTAIMMLLRIDYETRLEKAQAFVRGSR pbpb_ecoli Penicillin-binding protein 1B mrcB pbpF ponB b0149 JW0145 Murein polymerase protein protein ecoli Escherichia coli (strain K12) MAGNDREPIGRKGKPTRPVKQKVSRRRYEDDDDYDDYDDYEDEEPMPRKGKGKGKGRKPRGKRGWLWLLLKLAIVFAVLIAIYGVYLDQKIRSRIDGKVWQLPAAVYGRMVNLEPDMTISKNEMVKLLEATQYRQVSKMTRPGEFTVQANSIEMIRRPFDFPDSKEGQVRARLTFDGDHLATIVNMENNRQFGFFRLDPRLITMISSPNGEQRLFVPRSGFPDLLVDTLLATEDRHFYEHDGISLYSIGRAVLANLTAGRTVQGASTLTQQLVKNLFLSSERSYWRKANEAYMALIMDARYSKDRILELYMNEVYLGQSGDNEIRGFPLASLYYFGRPVEELSLDQQALLVGMVKGASIYNPWRNPKLALERRNLVLRLLQQQQIIDQELYDMLSARPLGVQPRGGVISPQPAFMQLVRQELQAKLGDKVKDLSGVKIFTTFDSVAQDAAEKAAVEGIPALKKQRKLSDLETAIVVVDRFSGEVRAMVGGSEPQFAGYNRAMQARRSIGSLAKPATYLTALSQPKIYRLNTWIADAPIALRQPNGQVWSPQNDDRRYSESGRVMLVDALTRSMNVPTVNLGMALGLPAVTETWIKLGVPKDQLHPVPAMLLGALNLTPIEVAQAFQTIASGGNRAPLSALRSVIAEDGKVLYQSFPQAERAVPAQAAYLTLWTMQQVVQRGTGRQLGAKYPNLHLAGKTGTTNNNVDTWFAGIDGSTVTITWVGRDNNQPTKLYGASGAMSIYQRYLANQTPTPLNLVPPEDIADMGVDYDGNFVCSGGMRILPVWTSDPQSLCQQSEMQQQPSGNPFDQSSQPQQQPQQQPAQQEQKDSDGVAGWIKDMFGSN ftsn_ecoli Cell division protein FtsN ftsN msgA b3933 JW3904 protein protein ecoli Escherichia coli (strain K12) MAQRDYVRRSQPAPSRRKKSTSRKKQRNLPAVSPAMVAIAAAVLVTFIGGLYFITHHKKEESETLQSQKVTGNGLPPKPEERWRYIKELESRQPGVRAPTEPSAGGEVKTPEQLTPEQRQLLEQMQADMRQQPTQLVEVPWNEQTPEQRQQTLQRQRQAQQLAEQQRLAQQSRTTEQSWQQQTRTSQAAPVQAQPRQSKPASSQQPYQDLLQTPAHTTAQSKPQQAAPVARAADAPKPTAEKKDERRWMVQCGSFRGAEQAETVRAQLAFEGFDSKITTNNGWNRVVIGPVKGKENADSTLNRLKMAGHTNCIRLAAGG ftsi-ftsw-14 1307172 n/a 1307175 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role neutral component neutral component P0ABG4:p.Pro375Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P0ABG4:p.Pro368Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4 ftsi-ftsw-16 1307172 n/a 1307176 unspecified role unspecified role neutral component neutral component t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307175 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation P0AD68:p.Arg23Cys mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Figure 4, Table 2 ftsw-mrcb-3 1307172 n/a 1307177 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Table 2 ftsw-mrcb-4 1307172 n/a 1307176 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation ha-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain false P0ABG4:p.Pro375Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P0ABG4:p.Pro368Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307177 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Table 2 ftsn-ftsw-5 1307172 n/a 1307178 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P0ABG4:p.Pro368Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false ha-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain false P0ABG4:p.Pro375Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Table 2 ftsn-ftsw-4 1307172 n/a 1307178 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role neutral component neutral component t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Table 2 ftsi-ftsw-13 1307172 n/a 1307175 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation t18-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation ha-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain false t25-(g4s)3 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P0ABG4:p.Pro375Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P0ABG4:p.Pro368Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Table 2 ftsi-ftsw-2 1307173 n/a 1307175 unspecified role unspecified role fluorescence donor fluorescence donor nucl transformation nucleic acid transformation nucl transformation mkusabira orange fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307176 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept nucl transformation nucleic acid transformation nucl transformation mcherry red fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2 ftsi-ftsw-9 1307173 n/a 1307175 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept nucl transformation nucleic acid transformation nucl transformation mcherry red mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1307176 unspecified role unspecified role fluorescence donor fluorescence donor nucl transformation nucleic acid transformation nucl transformation mkusabira orange fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figures 2 & 5 ftsi-ftsi 1307173 n/a 1307175 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept nucl transformation nucleic acid transformation nucl transformation region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false mcherry red fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1307175 unspecified role unspecified role fluorescence donor fluorescence donor nucl transformation nucleic acid transformation nucl transformation mkusabira orange fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1307244 1307240 proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2 & 5 ftsi-ftsw-3 1307174 n/a Claudine Fraipont 1307176 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation ha-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain false n/a Claudine Fraipont 1307175 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Figure 3