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/ leonardi-2003-1 Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex. ecobd Escherichia coli (strain B / BL21-DE3) anti tag coip anti tag coimmunoprecipitation anti tag coip protein sequence protein sequence identification protein sequence Leonardi R., Fairhurst S.A., Kriek M., Lowe D.J., Roach P.L. Three different plasmids were used: A pET plasmid encoding ThiGH-His was named pRL800. pET and pBAD plasmids encoding ThiFSGH-His were named pRL1000 and pRL1020, respectively. Eluted proteins were first visualized by SDS-PAGE before analysing their identity by N-terminal protein sequencing and electrospray mass spectrometry. plr2@soton.ac.uk FEBS Lett. (0014-5793) 2003 Only protein-protein interactions imex curation leonardi-2003-2 Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by staining molecular weight estimation by staining weight by staining Leonardi R., Fairhurst S.A., Kriek M., Lowe D.J., Roach P.L. Three different plasmids were used: A pET plasmid encoding ThiGH-His was named pRL800. pET and pBAD plasmids encoding ThiFSGH-His were named pRL1000 and pRL1020, respectively. Eluted proteins were first visualized by SDS-PAGE before analysing their identity by N-terminal protein sequencing and electrospray mass spectrometry. plr2@soton.ac.uk FEBS Lett. (0014-5793) 2003 Only protein-protein interactions imex curation thih_ecoli 2-iminoacetate synthase thiH b3990 JW3953 Tyrosine lyase Dehydroglycine synthase protein protein ecoli Escherichia coli (strain K12) MKTFSDRWRQLDWDDIRLRINGKTAADVERALNASQLTRDDMMALLSPAASGYLEQLAQRAQRLTRQRFGNTVSFYVPLYLSNLCANDCTYCGFSMSNRIKRKTLDEADIARESAAIREMGFEHLLLVTGEHQAKVGMDYFRRHLPALREQFSSLQMEVQPLAETEYAELKQLGLDGVMVYQETYHEATYARHHLKGKKQDFFWRLETPDRLGRAGIDKIGLGALIGLSDNWRVDSYMVAEHLLWLQQHYWQSRYSVSFPRLRPCTGGIEPASIMDERQLVQTICAFRLLAPEIELSLSTRESPWFRDRVIPLAINNVSAFSKTQPGGYADNHPELEQFSPHDDRRPEAVAAALTAQGLQPVWKDWDSYLGRASQRL thig_ecoli Thiazole synthase thiG b3991 JW5549 protein protein ecoli Escherichia coli (strain K12) MLRIADKTFDSHLFTGTGKFASSQLMVEAIRASGSQLVTLAMKRVDLRQHNDAILEPLIAAGVTLLPNTSGAKTAEEAIFAAHLAREALGTNWLKLEIHPDARWLLPDPIETLKAAETLVQQGFVVLPYCGADPVLCKRLEEVGCAAVMPLGAPIGSNQGLETRAMLEIIIQQATVPVVVDAGIGVPSHAAQALEMGADAVLVNTAIAVADDPVNMAKAFRLAVEAGLLARQSGPGSRSYFAHATSPLTGFLEASA thih-thig-1 1457734 n/a 1457736 unspecified role unspecified role bait bait c-terminal 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 1457737 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig. 2a The 26kD band was much stronger for the plasmids expressing ThiFSGH. thih-thig-2 1457735 n/a 1457737 unspecified role unspecified role prey prey n/a 1457736 unspecified role unspecified role bait bait c-terminal 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 direct interaction direct interaction false false false Fig. 2b The 26kD band was much stronger for the plasmids expressing ThiFSGH.