Crystallization (Comment) | Organism |
---|---|
FNR in complex with NADP+, two different crystal forms, mixing of 0.001 ml of 10 mg/ml protein and 2.5 mM NADP+ with 0.001 ml of reservoir solution containing 0.1 M HEPES buffer, pH 7.5, 30% 1,2-propanediol, and 20% PEG 400 for form I, and 20% PEG 3350, 0.2 M sodium fluoride, and 5% trehalose for form II, 20°C, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution, respetively, molecular replacement | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
H324F | site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type FNR | Bacillus subtilis |
H324S | site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type FNR | Bacillus subtilis |
additional information | site-directed mutagenesis of NADPH-specific BsFNR to replace Arg186, Asp187, Arg190, and His324 with the residues occurring in NADH/NADPH-bispecific Chlorobium tepidum FNR | Bacillus subtilis |
R186G | site-directed mutagenesis, replacement of Arg186 with glycine leads to drastically reduced amounts of recombinant protein | Bacillus subtilis |
R186G/D187H | site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type FNR | Bacillus subtilis |
R186G/D187H/R190Q | site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type FNR | Bacillus subtilis |
R190Q | site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type FNR | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 reduced ferredoxin + NADP+ | Bacillus subtilis | - |
2 oxidized ferredoxin + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | O05268 | gene yumC | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 reduced ferredoxin + NADP+ | - |
Bacillus subtilis | 2 oxidized ferredoxin + NADPH + H+ | - |
r | |
2 reduced ferredoxin + NADP+ | with K3[Fe(CN)6] as electron acceptor in the enzyme assay. Ferredoxin is a low-redox-potential iron-sulfur protein. BsFNR features two distinct binding domains for FAD and NADPH, the deduced mode of NADP+ binding to the BsFNR molecule is nonproductive in that the nicotinamide and isoalloxazine rings are over 15A A apart, binding structures, overview | Bacillus subtilis | 2 oxidized ferredoxin + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | structure, primary sequence and oligomeric conformation, comparisons, overview | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
ferredoxin-NADP+ oxidoreductase | - |
Bacillus subtilis |
FNR | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | BsFNR features two distinct binding domains for FAD and NADPH, binding structure, overview. A unique C-terminal extension covers the re-face of the isoalloxazine moiety of FAD. Tyr50 in the FAD-binding region and His324 in the Cterminal extension stack on the si- and re-faces of the isoalloxazine ring of FAD, respectively | Bacillus subtilis |