Literature summary for 2.7.7.2 extracted from

Herguedas, B.; Lans, I.; Sebastian, M.; Hermoso, J.A.; Martinez-Julvez, M.; Medina, M.
Structural insights into the synthesis of FMN in prokaryotic organisms (2015), Acta Crystallogr. Sect. D, 71, 2526-2542.

Cloned(Commentary)

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 (Commentary)

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

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

Metals/Ions	Comment	Organism	Structure
Ca2+	can substitute for Mg2+	Corynebacterium ammoniagenes
Mg2+	required	Corynebacterium ammoniagenes

Natural Substrates/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Corynebacterium ammoniagenes	Q59263	gene ribF	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant RFK module of enzyme FADS, DELTA(1-182)CaFADS, from Escherichia coli strain BL21(DE3)	Corynebacterium ammoniagenes

Substrates and Products (Substrate)

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

Subunits	Comment	Organism
hexamer	dimer-of-trimers organization with the catalytic sites of two modules of neighbouring protomers approaching each other	Corynebacterium ammoniagenes

Synonyms

Synonyms	Comment	Organism
FADS	-	Corynebacterium ammoniagenes
FMN:ATP adenylyltransferase	-	Corynebacterium ammoniagenes
FMNAT	-	Corynebacterium ammoniagenes

Cofactor

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

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