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

FEBS Letters

Bonaccorsi di Patti, Maria Carmela¹  Paronetto, Maria Paola¹  Dolci, Valeria¹  Felice, Maria Rosa²  Musci, Giovanni²  Lania, Amalia² 

[1] Department of Biochemical Sciences, University of Rome ‘La Sapienza’, Rome 00185, Italy;Department of Microbiological, Genetic and Molecular Sciences, University of Messina, Salita Sperone 31, S. Agata Messina 98166, Italy

关键词: Fet3;    Ferroxidase;    High affinity iron uptake;    Site-directed mutagenesis;   

DOI  :  10.1016/S0014-5793(01)03131-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 K _m. 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.

【 授权许可】

Unknown