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Literature summary extracted from

  • Adachi, M.; Taylor, A.; Hart, P.; Fitzpatrick, P.
    Mechanistic and structural analyses of the roles of active site residues in yeast polyamine oxidase Fms1: characterization of the N195A and D94N enzymes (2012), Biochemistry, 51, 8690-8697.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.3.17 recombinant expression of wild-type and mutant enzymes Saccharomyces cerevisiae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.3.17 purified recombinant enzyme mutant N195A, sitting drop vapour diffusion method, mixing of protein solution, containing 25 mM HEPES, 25 mM NaCl, and 2% glycerol, pH 7.5, in a 1:1 ratio with buffer containing 30 mM diethylene glycol, 30 mM triethylene glycol, 30 mM tetraethylene glycol, 30 mM pentaethylene glycol, 10% ethylene glycol, 20% PEG 8000, and 0.1 MES-imidazole, pH 6.5, 1 week, X-ray diffraction structure determination and analysis at 2.0 A resolution Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
1.5.3.17 D94N site-directed mutagenesis, the mutation primarily affects the reductive half-reaction. The mutant shows 20-40fold reduced rate constant for flavin reduction with spermine and 450fold with N1-acetylspermine compared to the wild-type enzyme Saccharomyces cerevisiae
1.5.3.17 N195A site-directed mutagenesis, the mutation primarily affects the reductive half-reaction. The mutant shows 20-40fold reduced rate constant for flavin reduction with spermine and 450fold with N1-acetylspermine compared to the wild-type enzyme. Mutant N195A enzyme shows structure with a molecule of tetraethylene glycol in the active site, the mutation has no effect on the protein structure Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.3.17 additional information
-
 additional information steady-state kinetics and rapid-reaction kinetics of wild-type and mutant enzymes, overview Saccharomyces cerevisiae
1.5.3.17 0.011
-
 N1-acetylspermine pH 9.0, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae
1.5.3.17 0.015
-
 O2 pH 9.0, 25°C, recombinant mutant N195A, with spermine Saccharomyces cerevisiae
1.5.3.17 0.02
-
 O2 pH 9.0, 25°C, recombinant mutant N195A, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 0.03
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant D94N Saccharomyces cerevisiae
1.5.3.17 0.0436
-
 O2 pH 9.0, 25°C, recombinant wild-type Fms1, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 0.091
-
 O2 pH 9.0, 25°C, recombinant wild-type Fms1, with spermine Saccharomyces cerevisiae
1.5.3.17 0.097
-
 O2 pH 9.0, 25°C, recombinant mutant D94N, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 0.104
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 0.118
-
 spermine pH 9.35, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae
1.5.3.17 0.127
-
 spermine pH 9.35, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 0.192
-
 O2 pH 9.0, 25°C, recombinant mutant D94N, with spermine Saccharomyces cerevisiae
1.5.3.17 0.32
-
 spermine pH 9.35, 25°C, recombinant mutant D94N Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.3.17 N1-acetylspermine + O2 + H2O Saccharomyces cerevisiae
-
 spermidine + N-acetyl-3-aminopropanal + H2O2
-
 ?
1.5.3.17 spermine + O2 + H2O Saccharomyces cerevisiae
-
 spermidine + 3-aminopropanal + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.3.17 Saccharomyces cerevisiae P50264
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.3.17 recombinant wild-type and mutant enzymes Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.3.17 N1-acetylspermine + O2 + H2O
-
 Saccharomyces cerevisiae spermidine + N-acetyl-3-aminopropanal + H2O2
-
 ?
1.5.3.17 spermine + O2 + H2O
-
 Saccharomyces cerevisiae spermidine + 3-aminopropanal + H2O2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.5.3.17 Fms1
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.3.17 25
-
 assay at Saccharomyces cerevisiae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.3.17 2
-
 spermine pH 9.35, 25°C, recombinant mutant D94N Saccharomyces cerevisiae
1.5.3.17 4.9
-
 spermine pH 9.35, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 8.1
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 9.7
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant D94N Saccharomyces cerevisiae
1.5.3.17 15.1
-
 N1-acetylspermine pH 9.0, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae
1.5.3.17 39
-
 spermine pH 9.35, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.3.17 9
-
 substrate N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 9.35
-
 substrate spermine Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.3.17 FAD dependent on Saccharomyces cerevisiae

General Information

EC Number General Information Comment Organism
1.5.3.17 additional information the active site of Fms1 contains three amino acid residues positioned to interact with the polyamine substrate, His67, Asn195, and Asp94. These three residues form a hydrogen-bonding triad with Asn195 being the central residue. His67 is important both for interacting with the substrate and for maintaining the hydrogen bonds in the triad Saccharomyces cerevisiae
1.5.3.17 physiological function flavoprotein Fms1 catalyzes the oxidation of spermine in the biosynthetic pathway for pantothenic acid Saccharomyces cerevisiae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5.3.17 4.1
-
 spermine pH 9.35, 25°C, recombinant mutant D94N Saccharomyces cerevisiae
1.5.3.17 38.5
-
 spermine pH 9.35, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 78
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant N195A Saccharomyces cerevisiae
1.5.3.17 100
-
 O2 pH 9.0, 25°C, recombinant mutant D94N, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 104
-
 O2 pH 9.0, 25°C, recombinant mutant D94N, with spermine Saccharomyces cerevisiae
1.5.3.17 320
-
 N1-acetylspermine pH 9.0, 25°C, recombinant mutant D94N Saccharomyces cerevisiae
1.5.3.17 327
-
 O2 pH 9.0, 25°C, recombinant mutant N195A, with spermine Saccharomyces cerevisiae
1.5.3.17 330
-
 spermine pH 9.35, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae
1.5.3.17 358
-
 O2 pH 9.0, 25°C, recombinant wild-type Fms1, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 405
-
 O2 pH 9.0, 25°C, recombinant mutant N195A, with N1-acetylspermine Saccharomyces cerevisiae
1.5.3.17 428
-
 O2 pH 9.0, 25°C, recombinant wild-type Fms1, with spermine Saccharomyces cerevisiae
1.5.3.17 1400
-
 N1-acetylspermine pH 9.0, 25°C, recombinant wild-type Fms1 Saccharomyces cerevisiae