Research article summary (published 27 Sep 2009):

Monitoring the iron status of the ferroxidase center of Escherichia coli bacterioferritin using fluorescence spectroscopy.

Full Abstract

Ferritins solubilize and detoxify the essential metal iron through formation of a ferric mineral within the protein's central cavity. Key to this activity is an intrasubunit catalytic dinuclear iron center called the ferroxidase center. Here we show that the fluorescence intensity of Escherichia coli bacterioferritin (BFR), due to the presence of two tryptophan residues (Trp35 and Trp133) in each of the 24 subunits, is highly sensitive to the iron status of the ferroxidase center and is quenched to different extents by Fe2+ and Fe3+. Recovery of the quench following oxidation of Fe2+ to Fe3+ at the ferroxidase center was not observed, indicating that the di-Fe3+ form of the center is stable. Studies of the single-tryptophan variants W35F and W133F showed that Trp133, which lies approximately 10 A from the ferroxidase center, is primarily responsible for the observed fluorescence sensitivity to iron, while studies of a stable E. coli BFR subunit dimer demonstrated that the observed quench properties are principally derived from the interaction of iron with tryptophan residues within the subunit dimer. A double-tryptophan variant (W35F/W133F) was found to exhibit fluorescence from the seven tyrosine residues present in each subunit, which was also sensitive to the iron status of the ferroxidase center. Finally, we demonstrate using Zn2+, a potent competitive inhibitor of Fe2+ binding and oxidation, that the fluorescence response can be used to monitor the loss of iron from the ferroxidase center.

Author information

Author/s: Lawson, Tamara L (TL); Crow, Allister (A); Lewin, Allison (A); Yasmin, Samina (S); Moore, Geoffrey R (GR); Le Brun, Nick E (NE);

Affiliation: Centre for Molecular and Structural Biochemistry, School of Chemistry, University of East Anglia, Norwich NR4 7TJ, UK.

Grants: BB/D001943/1 (Agency:Biotechnology and Biological Sciences Research Council) ; (Agency:Wellcome Trust)

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't

Journal: Biochemistry (Biochemistry), published in United States. (Language: eng)

Reference: 2009-Sep; vol 48 (issue 38) : pp 9031-9

Dates: Created 2009/09/22; Completed 2009/10/09;

PMID: 19705876, status: MEDLINE (last retrieval date: 10/9/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Amino Acid Substitution Bacterial Proteins - chemistry; genetics; metabolism Base Sequence Cytochrome b Group - chemistry; genetics; metabolism DNA Primers - genetics Dimerization Escherichia coli Proteins - chemistry; genetics; metabolism Ferritins - chemistry; genetics; metabolism

Iron - chemistry; metabolism Models, Molecular Mutagenesis, Site-Directed Oxidation-Reduction Protein Subunits Recombinant Proteins - chemistry; genetics; metabolism Spectrometry, Fluorescence Tryptophan - chemistry

Associated Chemicals: Bacterial Proteins (0) ; Cytochrome b Group (0) ; DNA Primers (0) ; Escherichia coli Proteins (0) ; Protein Subunits (0) ; Recombinant Proteins (0) ; Tryptophan (73-22-3) ; Iron (7439-89-6) ; Ferritins (9007-73-2) ; bacterioferritin (9035-38-5)

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