Purification and characterization of Fet3 protein, a yeast homologue of ceruloplasmin.

The Journal of Biological Chemistry
D M De SilvaJ Kaplan

Abstract

The FET3 gene product of Saccharomyces cerevisiae is an essential component of the high affinity iron transport system. Based on FET3 sequence homology to the multicopper oxidase family and iron oxidation studies in spheroplasts (De Silva, D. M., Askwith, C. C., Eide, D., and Kaplan, J. (1995) J. Biol. Chem. 270, 1098-1101), it was hypothesized that the Fet3 protein (Fet3p) was a cell surface ferroxidase. To further characterize the protein, we have isolated Fet3p from yeast membranes and purified the protein to apparent homogeneity. Consistent with its localization at the plasma membrane, Fet3p is a glycosylated protein. SDS-polyacrylamide gel electrophoresis analysis showed that the protein was present in two differentially glycosylated forms of approximately 120 and 100 kDa. Purified Fet3p is a copper-containing protein that is able to catalyze the oxidation of a variety of organic compounds in addition to ferrous iron. Azide and metal chelators strongly inhibited enzyme activity. Iron appeared to be the best substrate for the enzyme, and the apparent Km for ferrous oxidation was 2 microM. Interestingly, Fet3p was able to effectively catalyze the incorporation of iron onto apotransferrin. We conclude that Fet3p is a ferro-O2...Continue Reading

References

Mar 1, 1992·Archives of Biochemistry and Biophysics·D de SilvaS D Aust
May 1, 1990·Molecular and Cellular Biology·A DancisJ G Barriocanal
Jan 26, 1990·European Journal of Biochemistry·A Messerschmidt, R Huber
Oct 14, 1983·Biochemical and Biophysical Research Communications·R F Boyer, B E Schori
Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·Z L HarrisJ D Gitlin
Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·D S YuanR D Klausner
Jan 20, 1995·The Journal of Biological Chemistry·D M De SilvaJ Kaplan
Jan 26, 1996·The Journal of Biological Chemistry·G MusciL Calabrese
Aug 30, 1996·Journal of Molecular Biology·U RydeB O Roos

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Citations

Dec 22, 1999·Archives of Biochemistry and Biophysics·M C Bonaccorsi di PattiL Calabrese
Nov 16, 2007·European Biophysics Journal : EBJ·Ian G Macreadie
Aug 16, 2012·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Alexandra V SoldatovaThomas G Spiro
Jul 17, 1998·Current Opinion in Chemical Biology·D R WingeC Srinivasan
Oct 2, 2002·Molecular Microbiology·Wilhelmina M HustonAlastair G McEwan
Oct 3, 1999·European Journal of Biochemistry·G MusciL Calabrese
May 21, 2011·The Journal of Biological Chemistry·Doreen KochDietrich H Nies
Aug 30, 2001·Applied and Environmental Microbiology·C A FrancisB M Tebo
Aug 30, 2001·Applied and Environmental Microbiology·C A Francis, B M Tebo
Oct 9, 2003·Applied and Environmental Microbiology·Luis F LarrondoDaniel Cullen
Sep 10, 2005·Applied and Environmental Microbiology·S SitthisakR K Jayaswal
May 2, 2007·Infection and Immunity·John R Erb-Downward, Mairi C Noverr
Apr 25, 2000·Annual Review of Medicine·S Sheth, G M Brittenham
Mar 7, 2013·Genetics·Martha S Cyert, Caroline C Philpott
Apr 18, 2008·World Journal of Gastroenterology : WJG·Isabel SimonWolfgang Stremmel
Mar 5, 2014·Chemical Reviews·Edward I SolomonLi Tian
Apr 1, 2005·Proceedings of the National Academy of Sciences of the United States of America·Samuel M WebbBradley M Tebo
Nov 13, 1998·Proceedings of the National Academy of Sciences of the United States of America·S R Davis-KaplanJ Kaplan
Feb 27, 2003·Molecular Microbiology·Daniel J Kosman
Sep 15, 2006·Fungal Genetics and Biology : FG & B·Phil Kersten, Dan Cullen
Jun 28, 2003·FEMS Microbiology Reviews·Imke SchröderSimon de Vries
Feb 24, 2015·Insect Biochemistry and Molecular Biology·Zeyu PengMaureen J Gorman
Jul 27, 2002·Annual Review of Microbiology·Anthony Van HoJerry Kaplan
May 17, 2014·Biochemical and Biophysical Research Communications·Deepti SinghJugsharan Singh Virdi
Jul 10, 2016·Microbial Pathogenesis·Moiz A AnsariSaif Hameed
Jan 28, 2017·Metallomics : Integrated Biometal Science·Cristina N Butterfield, Bradley M Tebo
Aug 29, 2003·The Journal of Biological Chemistry·Xiaoli ShiZhiwu Zhu
Apr 10, 2003·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Tzu-Pin WangDaniel J Kosman
Feb 13, 1999·The Journal of Biological Chemistry·D Radisky, J Kaplan
Apr 29, 1998·The Journal of Biological Chemistry·R F HassettD J Kosman
Aug 26, 1998·The Journal of Biological Chemistry·C C Askwith, J Kaplan

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