The essential role of Glu‐185 and Tyr‐354 residues in the ferroxidase activity of Saccharomyces cerevisiae Fet3

FEBS Letters

Bonaccorsi di Patti, Maria Carmela²  Felice, Maria Rosa¹  Musci, Giovanni¹  Camuti, Angela Pia¹  Lania, Amalia¹ 

[1] Department of Organic and Biological Chemistry, University of Messina, Salita Sperone 31, S. Agata Messina 98166, Italy;Department of Biochemical Sciences, University of Rome La Sapienza, Rome 00185, Italy

关键词: Fet3;    Ferroxidase;    Iron homeostasis;    Site-directed mutagenesis;   

DOI  :  10.1016/S0014-5793(00)01435-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The structural determinants required for ferroxidase activity by the yeast multicopper oxidase Fet3 have been partially clarified by site-directed mutagenesis based on homology modeling. Glu-185 and Tyr-354 were substituted with Ala and Phe, respectively. Fet3 E185A retained ca. 5% residual ferroxidase catalytic efficiency, and almost 40% oxidase efficiency. On the other hand, Fet3 Y354F exhibited 50% residual efficiency as a ferroxidase and more than 70% as an oxidase. These results provide new insights in the mechanism of iron binding and oxidation by Fet3, establishing the essential role of Glu-185 and Tyr-354, and allowing to dissect ferroxidase from non-iron oxidase activity.

【 授权许可】

Unknown