Effect of Val 73 --> Trp mutation on the reaction of "cambialistic" superoxide dismutase from Propionibacterium shermanii with hydrogen peroxide.
The H2O2 inactivation of the "cambialistic" superoxide dismutases from Propionibacterium shermanii, which is active with either iron or manganese at the active site, has been studied in the native and Val 73 --> Trp mutant enzymes. The wild-type iron-containing form of this enzyme is much more resistant to treatment with H2O2 with respect to the other metal-specific Fe superoxide dismutase isoenzymes. After incubation with high amounts of H2O2 the enzyme maintains more than 40% of the initial activity. The activity of the Val 73 --> Trp mutant drastically decreases to less than 5% of the initial activity after incubation with hydrogen peroxide. Amino acid analysis of the H2O2-treated mutant enzyme evidenced the loss of the Trp 73 residue which is shown to play a critical role in the stabilization of the monomer fold of the enzyme. On the other hand, the manganese-containing wild-type and mutant enzymes were completely resistant toward H2O2 demonstrating the specific role of iron in the inactivation process.
Gabbianelli
R
R
INFM, Department of Biology, University of Rome "Tor Vergata,", Via della Ricerca Scientifica e Tecnologica, Rome, 00133, Italy.
Battistoni
A
A
Capo
C
C
Polticelli
F
F
Rotilio
G
G
Meier
B
B
Desideri
A
A
D001665:Binding Sites; D019281:Dimerization; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007501:Iron / Q000032:analysis; D008345:Manganese / Q000032:analysis; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011424:Propionibacterium / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014364:Tryptophan / Q000235:genetics*; D014633:Valine / Q000235:genetics*
D008345:Manganese; D014364:Tryptophan; D006861:Hydrogen Peroxide; D007501:Iron; D013482:Superoxide Dismutase; D014633:Valine