TY - JOUR
T1 - Mutational analysis of the iron binding site of Saccharomyces cerevisiae ferroxidase Fet3. An in vivo study
AU - Bonaccorsi di Patti, Maria Carmela
AU - Paronetto, Maria Paola
AU - Dolci, Valeria
AU - Felice, Maria Rosa
AU - Lania, Amalia
AU - Musci, Giovanni
PY - 2001/11/23
Y1 - 2001/11/23
N2 - The role of residues predicted to be involved in the binding of iron by the yeast ferroxidase Fet3 has been studied by site-directed mutagenesis. The effect of Fet3 mutations E185A, E185Q, Y354F, D409V and H489D has been investigated in vivo by kinetic analyses of high affinity iron uptake. Our results indicate that Glu-185 is critical for the binding of iron, since substitution of this residue with Ala or Gln strongly affects both growth and the kinetic parameters of high affinity iron uptake, greatly increasing Km. Mutations Y354F and D409V result in less severe alteration of high affinity iron uptake, while mutant H489D is unable to grow under conditions of iron limitation.
AB - The role of residues predicted to be involved in the binding of iron by the yeast ferroxidase Fet3 has been studied by site-directed mutagenesis. The effect of Fet3 mutations E185A, E185Q, Y354F, D409V and H489D has been investigated in vivo by kinetic analyses of high affinity iron uptake. Our results indicate that Glu-185 is critical for the binding of iron, since substitution of this residue with Ala or Gln strongly affects both growth and the kinetic parameters of high affinity iron uptake, greatly increasing Km. Mutations Y354F and D409V result in less severe alteration of high affinity iron uptake, while mutant H489D is unable to grow under conditions of iron limitation.
KW - Ferroxidase
KW - Fet3
KW - High affinity iron uptake
KW - Site-directed mutagenesis
UR - http://www.scopus.com/inward/record.url?scp=0035941079&partnerID=8YFLogxK
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U2 - 10.1016/S0014-5793(01)03131-3
DO - 10.1016/S0014-5793(01)03131-3
M3 - Article
C2 - 11728475
AN - SCOPUS:0035941079
VL - 508
SP - 475
EP - 478
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 3
ER -