Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme mutant H67Q, sitting drop vapor diffusion method, mixing of protein solution, in 25 mM HEPES, pH 7.5, in a 1:1 ratio with buffer containing 20% w/v PEG 3350, 0.2 M sodium acetate, and 0.1 M bis-Tris propane, pH 7.5, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
H67A | site-directed mutagenesis, the mutant shows a 2-3fold reduced first-order rate constant for flavin reduction and slightly altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
H67N | site-directed mutagenesis, the mutant shows a 2-3fold reduced first-order rate constant for flavin reduction and slightly altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
H67Q | site-directed mutagenesis, the mutant shows a 2-3fold reduced first-order rate constant for flavin reduction and slightly altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | seady-state kinetics and rapid-reaction kinetics of wild-type and mutant enzymes, overview | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Saccharomyces cerevisiae | flavoprotein oxidase Fms1 catalyzes the oxidation of spermine and N1-acetylspermine to spermidine and 3-aminopropanal or N-acetyl-3-aminopropanal | ? | - |
? | |
N1-acetylspermine + O2 + H2O | Saccharomyces cerevisiae | - |
spermidine + N-acetyl-3-aminopropanal + H2O2 | - |
? | |
spermine + O2 + H2O | Saccharomyces cerevisiae | - |
spermidine + 3-aminopropanal + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P50264 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes by nickel affinity chhromatography | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | flavoprotein oxidase Fms1 catalyzes the oxidation of spermine and N1-acetylspermine to spermidine and 3-aminopropanal or N-acetyl-3-aminopropanal | Saccharomyces cerevisiae | ? | - |
? | |
N1-acetylspermine + O2 + H2O | - |
Saccharomyces cerevisiae | spermidine + N-acetyl-3-aminopropanal + H2O2 | - |
? | |
spermine + O2 + H2O | - |
Saccharomyces cerevisiae | spermidine + 3-aminopropanal + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Fms1 | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
substrate N1-acetylspermine | Saccharomyces cerevisiae |
9.35 | - |
substrate spermine | Saccharomyces cerevisiae |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH profile for the reductive half-reaction of Fms1, a moiety with a pKa value of 7.2-8.3 must be unprotonated for amine oxidation, overview | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
additional information | the overall catalytic reactions of flavoprotein oxidases such as Fms1 can be divided into reductive and oxidative half-reactions. In the reductive half-reaction, binding of the oxidized substrate is followed by transfer of a hydride equivalent to the flavin to form reduced flavin and oxidized substrate. In the oxidative half-reaction, the reduced flavin is oxidized by molecular oxygen to form H2O2, the oxidized amine then dissociates from the enzyme. A moiety with a pKa value of 7.2-8.3 must be unprotonated for amine oxidation | Saccharomyces cerevisiae |