Application | Comment | Organism |
---|---|---|
drug development | HsRFK parameters differ from those of the so far evaluated bacterial counterparts, suggesting species-specific mechanisms in RFK catalysis. Thus, HsRFK is a potential therapeutic target because of its key functions, while bacterial RFK modules are potential antimicrobial targets | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
gene RFK, recombinant expression of His-tagged enzyme from codon-optimzed gene in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
analysis of the crystallographic structure of HsRFK in complex with FMN and ADP in either the open (PDB ID 1P4M) or the closed conformation (PDB ID 1Q9S) of the flavin binding site overview | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ADP | product inhibition, fitted to the Lineweaver-Burk equation for competitive inhibition | Homo sapiens | |
FMN | product inhibition, fitted to the Lineweaver-Burk equation for competitive inhibition | Homo sapiens | |
additional information | the inhibition mechanism of the products of the RFK reaction, FMN and ADP, kinetics, overview | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics and modeling, cooperativity model, pre-steady-state kinetics/pre-steady-state stopped-flow kinetics and dissociation constants, overview. Isothermal titration calorimetry, and Gibbs free energy flow for the interaction of HsRFK with substrates and products. Thermodynamics modulates the ligand binding landscape of HsRFK. Cooperativity coefficients, ANP and FLV ligands cooperate in their binding to HsRFK | Homo sapiens | |
0.0025 | - |
riboflavin | pH 7.0, 25°C, recombinant enzyme | Homo sapiens | |
0.03 | - |
ATP | pH 7.0, 25°C, recombinant enzyme | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + riboflavin | Homo sapiens | - |
ADP + FMN | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q969G6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme RFK from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage by a protease, followed by ammonium sulfate fractionation, hydrophobic interaction chromatography, and dialysis. HsRFK is purified free of flavin and adenine ligands | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + riboflavin | - |
Homo sapiens | ADP + FMN | - |
? | |
additional information | substrate binding structure analysis using crystal structures of substrate-bound RFK. Binding of HsRFK substrates is the kinetically preferred process compared to product inhibition, kinetic modeling, overview | Homo sapiens | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
More | structure-function analysis, apo-HsRFK structural model | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
HsRFK | - |
Homo sapiens |
RFK | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.7 | - |
ATP | pH 7.0, 25°C, recombinant enzyme | Homo sapiens | |
1.7 | - |
riboflavin | pH 7.0, 25°C, recombinant enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics, recombinant enzyme, overview | Homo sapiens | |
0.0025 | - |
FMN | pH 7.0, 25°C, recombinant enzyme | Homo sapiens | |
0.033 | - |
ADP | pH 7.0, 25°C, recombinant enzyme | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | different organisms, different regulatory strategies of RFKs, overview | Homo sapiens |
malfunction | enzyme RFK downregulation alters expression profiles of clock-controlled metabolic-genes and destroys flavins protection on stroke treatments, while its activity reduction links to protein-energy malnutrition and thyroid hormones decrease | Homo sapiens |
additional information | molecular dynamics simulations, structure-function analysis | Homo sapiens |
physiological function | human riboflavin kinase is an essential enzyme that catalyzes the biosynthesis of the flavin mononucleotide (FMN) cofactor using riboflavin (RF, vitamin B2) and ATP as substrates. Human riboflavin kinase (HsRFK) catalyzes vitamin B2 (riboflavin) phosphorylation to flavin mononucleotide (FMN), obligatory step in flavin cofactor synthesis. HsRFK expression is related to protection from oxidative stress, amyloid-beta toxicity, and some malignant cancers progression. HsRFK is also predicted as involved in a protein-protein association network that at the system level affects to different cellular processes | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
56.67 | - |
ATP | pH 7.0, 25°C, recombinant enzyme | Homo sapiens | |
680 | - |
riboflavin | pH 7.0, 25°C, recombinant enzyme | Homo sapiens |