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/ martinez_ju-1998-2 Lys75 of Anabaena ferredoxin-NADP+ reductase is a critical residue for binding ferredoxin and flavodoxin during electron transfer. in vitro In vitro oxidoreduct assay oxidoreductase assay oxidoreduct assay predetermined predetermined participant predetermined Martinez-Julvez M., Medina M., Hurley JK., Hafezi R., Brodie TB., Tollin G., Gomez-Moreno C. Cyanobacteria - Interaction dataset based on Cyanobacteria proteins and related species Electron transfer detected by NADPH-Dependant Cytochrome c Reductase activity. The steady state catalytic behaviours of the different FNR mutants were determined. Table 3 gomezm@pota.unizar.es Biochemistry (0006-2960) 1998 IMEx Only protein-protein interactions imex curation imex curation martinez_ju-1998-1 Lys75 of Anabaena ferredoxin-NADP+ reductase is a critical residue for binding ferredoxin and flavodoxin during electron transfer. in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Martinez-Julvez M., Medina M., Hurley JK., Hafezi R., Brodie TB., Tollin G., Gomez-Moreno C. Cyanobacteria - Interaction dataset based on Cyanobacteria proteins and related species Table 3 gomezm@pota.unizar.es Biochemistry (0006-2960) 1998 IMEx Only protein-protein interactions imex curation imex curation flav_nosso Flavodoxin isiB protein protein anaso Anabaena sp. (strain PCC 7119) MSKKIGLFYGTQTGKTESVAEIIRDEFGNDVVTLHDVSQAEVTDLNDYQYLIIGCPTWNIGELQSDWEGLYSELDDVDFNGKLVAYFGTGDQIGYADNFQDAIGILEEKISQRGGKTVGYWSTDGYDFNDSKALRNGKFVGLALDEDNQSDLTDDRIKSWVAQLKSEFGL fenr_nosso Ferredoxin--NADP reductase petH protein protein anaso Anabaena sp. (strain PCC 7119) MSNQGAFDGAANVESGSRVFVYEVVGMRQNEETDQTNYPIRKSGSVFIRVPYNRMNQEMQRITRLGGKIVTIQTVSALQQLNGRTTIATVTDASSEIAKSEGNGKATPVKTDSGAKAFAKPPAEEQLKKKDNKGNTMTQAKAKHADVPVNLYRPNAPFIGKVISNEPLVKEGGIGIVQHIKFDLTGGNLKYIEGQSIGIIPPGVDKNGKPEKLRLYSIASTRHGDDVDDKTISLCVRQLEYKHPESGETVYGVCSTYLTHIEPGSEVKITGPVGKEMLLPDDPEANVIMLATGTGIAPMRTYLWRMFKDAERAANPEYQFKGFSWLVFGVPTTPNILYKEELEEIQQKYPDNFRLTYAISREQKNPQGGRMYIQDRVAEHADELWQLIKNQKTHTYICGLRGMEEGIDAALSAAAAKEGVTWSDYQKDLKKAGRWHVETY fer1_nosso Ferredoxin-1 Ferredoxin I petF fdxV protein protein anaso Anabaena sp. (strain PCC 7119) MATFKVTLINEAEGTKHEIEVPDDEYILDAAEEQGYDLPFSCRAGACSTCAGKLVSGTVDQSDQSFLDDDQIEAGYVLTCVAYPTSDVVIQTHKEEDLY peth-isib-1 Steady state kinetics: flavodoxin (fld) and FNR 208035 N/A 208037 unspecified role unspecified role neutral component neutral component N/A 208038 unspecified role unspecified role neutral component neutral component P21890:p.Lys206Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) redox reaction oxidoreductase activity electron transfer reaction redox reaction false false false Fig. 5 All the kinetics parameters were obtained by fitting the data to the equation for a ping-pong mechanism. The steady state parameters are: kcat= 23 s-1, Km = 33.10-6 M. For the mutants Lys75Gln and Lys75Ser of FNR, the kcat are respectively 27 s-1 and 50 s-1. For the mutants Lys75Gln and Lys75Ser of FNR, The Km are respectively 632.10-6 M and 800.10-6 M. No reaction was observed for Lys75Glu. peth-petf-3 Steady state kinetics: ferredoxin (fd) and FNR 208035 N/A 208038 unspecified role unspecified role neutral component neutral component P21890:p.Lys206Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 208039 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association false false false All the kinetics parameters were obtained by fitting the data to the equation for a ping-pong mechanism. The steady state parameters are: kcat= 200 s-1, Km = 11.10-6 M. For the mutants Lys75Gln and Lys75Ser of FNR, the kcat are respectively 190 s-1 and 200 s-1. The Km could not be measured. No reaction was observed for Lys75Glu FNR. peth-petf-1 Differential spectroscopy : interaction between ferredoxin (fd) and FNR 208036 N/A 208039 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) N/A 208038 unspecified role unspecified role neutral component neutral component P21890:p.Lys206Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P21890:p.Lys206Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Mutations of the positively charged lysine 75 of FNR into neutral amino acid lys75Gln or lys75Ser increase the Kd respectively to Kd = 3.8.10-4 and Kd = 2.10-4. For the mutation of Lysine 75 into a negatively charged amino-acid Lys75Glu, the Kd could not be determined as no binding was detected. Both proteins are in oxidized form.