Protein Variants | Comment | Organism |
---|---|---|
D168A | site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme | [Candida] glabrata |
D181A | site-directed mutagenesis, the mutant shows reduced sensitivity to inhibition by FAD compared to the wild-type enzyme and has a much faster turnover rate than the wild-type enzyme | [Candida] glabrata |
D66A | site-directed mutagenesis, inactive mutant | [Candida] glabrata |
N62A | site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme | [Candida] glabrata |
N62S | site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme | [Candida] glabrata |
R297A | site-directed mutagenesis, the mutant shows a 2fold increased activity compared to the wild-type enzyme | [Candida] glabrata |
R297A/R300A | site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme | [Candida] glabrata |
R300A | site-directed mutagenesis, the mutant shows 93% reduced activity compared to the wild-type enzyme | [Candida] glabrata |
W184A | site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme | [Candida] glabrata |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
FAD | product inhibition, wild-type enzyme FMNAT is strongly inhibited by FAD, whereas D181A mutant enzyme has an attenuated product inhibition | [Candida] glabrata |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis of wild-type and mutant enzymes. The enzyme from Candida glabrata apparently binds its substrates with high affinity, but the overall turnover rate is very slow due to product inhibition | [Candida] glabrata | |
0.0005 | - |
FMN | pH and temperature not specified in the publication, mutant N62A | [Candida] glabrata | |
0.001 | - |
FMN | pH and temperature not specified in the publication, wild-type enzyme | [Candida] glabrata | |
0.0011 | - |
FMN | pH and temperature not specified in the publication, mutant N62S | [Candida] glabrata | |
0.0015 | - |
FMN | pH and temperature not specified in the publication, mutant D181A | [Candida] glabrata | |
0.0023 | - |
FMN | pH and temperature not specified in the publication, mutant R300A | [Candida] glabrata | |
0.003 | - |
FMN | pH and temperature not specified in the publication, mutant R297A | [Candida] glabrata | |
0.0071 | - |
FMN | pH and temperature not specified in the publication, mutant D168A | [Candida] glabrata | |
0.0093 | - |
FMN | pH and temperature not specified in the publication, mutant R297A/R300A | [Candida] glabrata | |
0.1994 | - |
FMN | pH and temperature not specified in the publication, mutant W184A | [Candida] glabrata |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, the interaction with the two Mg2+ ion coordinating waters by Asp168 is important for the catalytic activity of the enzym. Residues Asn62, Asp66, Asp168, and Arg297 interact either with ATP phosphate groups, or coordinate the catalytic Mg2+ ion either directly or indirectly through water moleculese | [Candida] glabrata |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | [Candida] glabrata | - |
diphosphate + FAD | - |
? | |
additional information | [Candida] glabrata | 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 |
---|---|---|---|
[Candida] glabrata | Q6FNA9 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + FMN = diphosphate + FAD | product release may be the rate-limiting step of the reaction | [Candida] glabrata |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | - |
[Candida] glabrata | 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 | [Candida] glabrata | ? | - |
? | |
additional information | the flavin motif is involved in flavin ligand binding, role of active site residues in the catalytic mechanism, overview. The isoalloxazine ring is sandwiched between the indole ring of Trp184 and the planar guanidinium group of Arg189, while the hydrophilic pyrimidine ring forms two specific hydrogen bonds between its C4 carbonyl and the main chain amide of Asp181, and between its N3 amide and the side chain of Asp181, respectively. Residues Asn62, Asp66, Asp168, and Arg297 interact either with ATP phosphate groups, or to coordinate the catalytic Mg2+ ion either directly or indirectly through water molecules. Arg297 might be involved in the interaction with the phosphate groups of both substrates, and helps in their positioning for the nucleophilic attack in the adenylyltransfer reaction | [Candida] glabrata | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme contains a core domain with a modified Rossman-fold topology and a C-terminal extension. The substrate binding and catalytic site is located at the interface of the two domains | [Candida] glabrata |
Synonyms | Comment | Organism |
---|---|---|
FMN adenylyltransferase | - |
[Candida] glabrata |
FMNAT | - |
[Candida] glabrata |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
FMN | pH and temperature not specified in the publication, mutant R300A | [Candida] glabrata | |
0.012 | - |
FMN | pH and temperature not specified in the publication, mutant R297A/R300A | [Candida] glabrata | |
0.017 | - |
FMN | pH and temperature not specified in the publication, mutant D168A | [Candida] glabrata | |
0.017 | - |
FMN | pH and temperature not specified in the publication, mutant N62S | [Candida] glabrata | |
0.042 | - |
FMN | pH and temperature not specified in the publication, mutant N62A | [Candida] glabrata | |
0.089 | - |
FMN | pH and temperature not specified in the publication, wild-type enzyme | [Candida] glabrata | |
0.21 | - |
FMN | pH and temperature not specified in the publication, mutant R297A | [Candida] glabrata | |
0.26 | - |
FMN | pH and temperature not specified in the publication, mutant W184A | [Candida] glabrata | |
0.88 | - |
FMN | pH and temperature not specified in the publication, mutant D181A | [Candida] glabrata |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | residues Asn62, Asp66, Asp168, and Arg297 interact either with ATP phosphate groups, or coordinate the catalytic Mg2+ ion either directly or indirectly through water moleculese | [Candida] glabrata |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
FAD | versus ATP, pH and temperature not specified in the publication, wild-type enzyme | [Candida] glabrata | |
0.012 | - |
FAD | versus FMN, pH and temperature not specified in the publication, wild-type enzyme | [Candida] glabrata |
General Information | Comment | Organism |
---|---|---|
evolution | eukaryotic FMNAT is related to phosphoadenosine phosphosulfate (PAPS) reductase family proteins and contains a core domain with a modified Rossman-fold topology and a C-terminal extension | [Candida] glabrata |
physiological function | flavocoenzymes, including flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are versatile redox cofactors involved in many fundamental cellular processes in all living organisms. FAD is synthesized from riboflavin obtained from the diet via two enzymatic steps catalyzed by riboflavin kinase (RFK, EC 2.7.1.26) and essential FMN adenylyltransferase (FMNAT,EC 2.7.7.2). Phosphorylation of riboflavin by RFK is crucial for specific absorption of the vitamin and is the physiologically rate-limiting step in the biosynthesis of flavocoenzymes, whereas product (FAD) feedback inhibition is observed for mammalian FMNAT, suggesting that biosynthesis of FAD is also regulated at the FMNAT reaction step | [Candida] glabrata |