UniProt UniProt http://www.uniprot.org banzhaf-2012-5 in vitro In vitro fluorescence fluorescence technology fluorescence tag fluorescence tag visualisation by fluorescence tag fluorescence Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W., Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W. w.vollmer@ncl.ac.uk Molecular microbiology 2012 Only protein-protein interactions imex curation imex curation banzhaf-2012-1 in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom western blot western blot Immuno blot Immuno blot Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W., Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W. w.vollmer@ncl.ac.uk Molecular microbiology 2012 Only protein-protein interactions imex curation imex curation banzhaf-2012-2 ecoli Escherichia coli (strain K12) adenylate cyclase adenylate cyclase complementation bacterial two-hybrid bacterial two-hybrid adenylate cyclase predetermined predetermined participant predetermined Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W., Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W. w.vollmer@ncl.ac.uk Molecular microbiology 2012 Only protein-protein interactions imex curation imex curation banzhaf-2012-3 ecoli Escherichia coli (strain K12) crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W., Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W. Crosslinking with 3,3'-Dithiobis(sulfosuccinimidylpropionate) (DTSSP) followed by immunoprecipitation with anti‐PBP2. w.vollmer@ncl.ac.uk Molecular microbiology 2012 Only protein-protein interactions imex curation imex curation banzhaf-2012-4 in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting tag visualisation tag visualisation Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W., Banzhaf M., van den Berg van Saparoea B., Terrak M., Fraipont C., Egan A., Philippe J., Zapun A., Breukink E., Nguyen-Distèche M., den Blaauwen T., Vollmer W. w.vollmer@ncl.ac.uk Molecular microbiology 2012 Only protein-protein interactions imex curation imex curation penicillin penicillin Penicillin penicillins penicillins small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mrda_ecoli Peptidoglycan D,D-transpeptidase MrdA mrdA pbpA b0635 JW0630 Penicillin-binding protein 2 protein protein ecoli Escherichia coli (strain K12) MKLQNSFRDYTAESALFVRRALVAFLGILLLTGVLIANLYNLQIVRFTDYQTRSNENRIKLVPIAPSRGIIYDRNGIPLALNRTIYQIEMMPEKVDNVQQTLDALRSVVDLTDDDIAAFRKERARSHRFTSIPVKTNLTEVQVARFAVNQYRFPGVEVKGYKRRYYPYGSALTHVIGYVSKINDKDVERLNNDGKLANYAATHDIGKLGIERYYEDVLHGQTGYEEVEVNNRGRVIRQLKEVPPQAGHDIYLTLDLKLQQYIETLLAGSRAAVVVTDPRTGGVLALVSTPSYDPNLFVDGISSKDYSALLNDPNTPLVNRATQGVYPPASTVKPYVAVSALSAGVITRNTTLFDPGWWQLPGSEKRYRDWKKWGHGRLNVTRSLEESADTFFYQVAYDMGIDRLSEWMGKFGYGHYTGIDLAEERSGNMPTREWKQKRFKKPWYQGDTIPVGIGQGYWTATPIQMSKALMILINDGIVKVPHLLMSTAEDGKQVPWVQPHEPPVGDIHSGYWELAKDGMYGVANRPNGTAHKYFASAPYKIAAKSGTAQVFGLKANETYNAHKIAERLRDHKLMTAFAPYNNPQVAVAMILENGGAGPAVGTLMRQILDHIMLGDNNTDLPAENPAVAAAEDH pbpa_ecoli Penicillin-binding protein 1A mrcA ponA b3396 JW3359 protein protein ecoli Escherichia coli (strain K12) MKFVKYFLILAVCCILLGAGSIYGLYRYIEPQLPDVATLKDVRLQIPMQIYSADGELIAQYGEKRRIPVTLDQIPPEMVKAFIATEDSRFYEHHGVDPVGIFRAASVALFSGHASQGASTITQQLARNFFLSPERTLMRKIKEVFLAIRIEQLLTKDEILELYLNKIYLGYRAYGVGAAAQVYFGKTVDQLTLNEMAVIAGLPKAPSTFNPLYSMDRAVARRNVVLSRMLDEGYITQQQFDQTRTEAINANYHAPEIAFSAPYLSEMVRQEMYNRYGESAYEDGYRIYTTITRKVQQAAQQAVRNNVLDYDMRHGYRGPANVLWKVGESAWDNNKITDTLKALPTYGPLLPAAVTSANPQQATAMLADGSTVALSMEGVRWARPYRSDTQQGPTPRKVTDVLQTGQQIWVRQVGDAWWLAQVPEVNSALVSINPQNGAVMALVGGFDFNQSKFNRATQALRQVGSNIKPFLYTAAMDKGLTLASMLNDVPISRWDASAGSDWQPKNSPPQYAGPIRLRQGLGQSKNVVMVRAMRAMGVDYAAEYLQRFGFPAQNIVHTESLALGSASFTPMQVARGYAVMANGGFLVDPWFISKIENDQGGVIFEAKPKVACPECDIPVIYGDTQKSNVLENNDVEDVAISREQQNVSVPMPQLEQANQALVAKTGAQEYAPHVINTPLAFLIKSALNTNIFGEPGWQGTGWRAGRDLQRRDIGGKTGTTNSSKDAWFSGYGPGVVTSVWIGFDDHRRNLGHTTASGAIKDQISGYEGGAKSAQPAWDAYMKAVLEGVPEQPLTPPPGIVTVNIDRSTGQLANGGNSREEYFIEGTQPTQQAVHEVGTTIIDNGEAQELF cefsulodin cefsulodin Cefsulodin cefsulodin cefsulodine cefsulodino cefsulodinum (6R,7R)-3-[(4-carbamoylpyridinium-1-yl)methyl]-8-oxo-7-{[(2R)-2-phenyl-2-sulfoacetyl]amino}-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 7beta-{[(2R)-2-phenyl-2-sulfoacetyl]amino}-3-(4-carbamoylpyridinium-1-yl)methyl-3,4-didehydrocepham-4-carboxylate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mecillinam mecillinam (2S,5R,6R)-6-[(azepan-1-ylmethylidene)amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid amdinocillin Coactin mecilinamo mecillinam mecillinamum penicillin HX 6beta-{[(azepan-1-yl)methylidene]amino}-2,2-dimethylpenam-3alpha-carboxylic acid small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ampicillin ampicillin aminobenzylpenicillin AMP ampicilina (2S,5R,6R)-6-{[(2R)-2-amino-2-phenylacetyl]amino}-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid (2S,5R,6R)-6-{[(2R)-2-AMINO-2-PHENYLETHANOYL]AMINO}-3,3-DIMETHYL-7-OXO-4-THIA-1-AZABICYCLO[3.2.0]HEPTANE-2-CARBOXYLIC ACID (2S,6R)-6-{[(2R)-2-amino-2-phenylethanoyl]amino}-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid 6-(D-(2-amino-2-phenylacetamido))-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo(3.2.0)heptane-2-carboxylic acid ABPC Ampicillin ampicillin ampicillin acid ampicillin anhydrous ampicilline ampicillinum Anhydrous ampicillin AP D-(-)-6-(alpha-aminophenylacetamido)penicillanic acid D-(-)-ampicillin 6beta-[(2R)-2-amino-2-phenylacetamido]-2,2-dimethylpenam-3alpha-carboxylic acid small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mrca-mrda-2 3727001 N/A 3727006 unspecified role unspecified role neutral component neutral component region bodipy label bodipy label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3727007 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3727008 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 6A cefsulodin-mrca-1 3727001 N/A 3727006 unspecified role unspecified role neutral component neutral component region bodipy label bodipy label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3727009 unspecified role unspecified role neutral component neutral component N/A 3727007 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 6A Cefsulodin specifically inhibits activity of PBP1A mecillinam-mrca-1 3727001 N/A 3727006 unspecified role unspecified role neutral component neutral component region bodipy label bodipy label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3727010 unspecified role unspecified role neutral component neutral component N/A 3727008 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 6A Mecillinam specifically inhibits activity of PBP2. mrda-mrca-1 3727002 N/A 3727008 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3727007 unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1B mrca-mrda-1 3727002 N/A 3727008 unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) N/A 3727007 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1B mrda-mrca-2 3727003 N/A 3727008 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed ecoli Escherichia coli (strain K12) N/A 3727007 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1C mrda-mrca-3 3727004 N/A 3727008 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3727007 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1D ampicillin-mrca-1 3727005 N/A 3727008 unspecified role unspecified role prey prey purified purified n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 3727011 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Fig. 1A