Cloned (Comment) | Organism |
---|---|
gene ribF, individual expression of the riboflavin kinase, RFK, module of enzyme FAD synthetase, FADS, i.e. DELTA(1-182)CaFADS, in Escherichia coli strain BL21(DE3) | Corynebacterium ammoniagenes |
Crystallization (Comment) | Organism |
---|---|
purified recombinant DELTA(1-182)CaFADS module in binary complex with ADP-Ca2+ and in ternary complex with FMN-ADP-Mg2+, mixing of 0.002 ml of 7.5-10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM MgCl2, 1mM FMN and/or 1 mM ADP, with 0.002 ml of reservoir solution containing 10-14% PEG 8000, 20% glycerol, 0.1 M MES-NaOH pH 6.5, 200 mM CaCl2 for the binary complex, or with 0.002 ml of reservoir solution containing 26-30% PEG 4000, 200 mM Li2SO4, 100 mM sodium acetate, pH 5.0, as well as 0.002 ml of 1 M NaI solution, for the ternary complex, X-ray diffraction structure determination and analysis at 1.65-2.15 A resolution, modelling | Corynebacterium ammoniagenes |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | can substitute for Mg2+ | Corynebacterium ammoniagenes | |
Mg2+ | required | Corynebacterium ammoniagenes |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | Corynebacterium ammoniagenes | - |
diphosphate + FAD | - |
? | |
additional information | Corynebacterium ammoniagenes | bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium ammoniagenes | Q59263 | gene ribF | - |
Purification (Comment) | Organism |
---|---|
recombinant RFK module of enzyme FADS, DELTA(1-182)CaFADS, from Escherichia coli strain BL21(DE3) | Corynebacterium ammoniagenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | - |
Corynebacterium ammoniagenes | diphosphate + FAD | - |
? | |
additional information | bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26 | Corynebacterium ammoniagenes | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | dimer-of-trimers organization with the catalytic sites of two modules of neighbouring protomers approaching each other | Corynebacterium ammoniagenes |
Synonyms | Comment | Organism |
---|---|---|
FADS | - |
Corynebacterium ammoniagenes |
FMN:ATP adenylyltransferase | - |
Corynebacterium ammoniagenes |
FMNAT | - |
Corynebacterium ammoniagenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | the ADP/ATP-binding pocket is closed in the ligand-free structure, structure overview. The ADP molecule establishes hydrogen bonds to residues located in L1c-FlapI: Gly196, Gly198 and Gly201 interact with the alpha- and beta-phosphates of ADP, whereas Pro207 and Thr208 (both belonging to the PTAN motif) form several hydrogen bonds via side-chain and main-chain atoms | Corynebacterium ammoniagenes |
General Information | Comment | Organism |
---|---|---|
evolution | whereas the N-terminal module of FADS lacks structural homology to eukaryotic FMNATs, the kinase module is homologous to monofunctional RFKs | Corynebacterium ammoniagenes |
additional information | molecular dynamics simulations of riboflavin kinase domain bound to FMN, ADP, and Mg2+, structure-function analysis, flavin-binding site structure in the RFK module of CaFADS, overview | Corynebacterium ammoniagenes |
physiological function | the essential cofactors of flavoproteins and flavoenzymes, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are synthesized from riboflavin in two sequential reactions: riboflavin phosphorylation is catalysed by an ATP-riboflavin kinase (RFK, EC 2.7.1.26) to produce FMN, which can be then converted to FAD by an FMN:ATP adenylyltranferase (FMNAT, EC 2.7.7.2). Bacteria contain a single bifunctional polypeptide called FAD synthetase (FADS) | Corynebacterium ammoniagenes |