Mutational analysis of the iron binding site of Saccharomyces cerevisiae ferroxidase Fet3. An in vivo study

Maria Carmela Bonaccorsi di Patti, Maria Paola Paronetto, Valeria Dolci, Maria Rosa Felice, Amalia Lania, Giovanni Musci

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)475-478
Number of pages4
JournalFEBS Letters
Volume508
Issue number3
DOIs
Publication statusPublished - Nov 23 2001

Keywords

  • Ferroxidase
  • Fet3
  • High affinity iron uptake
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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