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 mere 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.
The essential role of Glu185 and Tyr354 residues in the ferroxidase activity of Saccharomyces cerevisiae Fet3 / BONACCORSI DI PATTI, Maria Carmela; Felice, M. R.; Camuti, A. P.; Lania, A.; Musci, G.. - In: FEBS LETTERS. - ISSN 0014-5793. - 472:(2000), pp. 283-286.
The essential role of Glu185 and Tyr354 residues in the ferroxidase activity of Saccharomyces cerevisiae Fet3
BONACCORSI DI PATTI, Maria Carmela;
2000
Abstract
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 mere 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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
