Document Detail


Ferritin reactions: direct identification of the site for the diferric peroxide reaction intermediate.
MedLine Citation:
PMID:  15166287     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Ferritins managing iron-oxygen biochemistry in animals, plants, and microorganisms belong to the diiron carboxylate protein family and concentrate iron as ferric oxide approximately 10(14) times above the ferric K(s). Ferritin iron (up to 4,500 atoms), used for iron cofactors and heme, or to trap DNA-damaging oxidants in microorganisms, is concentrated in the protein nanocage cavity (5-8 nm) formed during assembly of polypeptide subunits, 24 in maxiferritins and 12 in miniferritins/DNA protection during starvation proteins. Direct identification of ferritin ferroxidase (F(ox)) sites, complicated by multiple types of iron-ferritin interactions, is now achieved with chimeric proteins where putative F(ox) site residues were introduced singly and cumulatively into an inactive host, an L maxiferritin. A dimagnesium ferritin cocrystal model guided site design and the diferric peroxo F(ox) intermediates (A at 650 nm) monitored activity. Diferric peroxo formation in chimeric and WT proteins had similar K(app) values and Hill coefficients. Catalytic activity required cooperative ferrous substrate binding to two sites A (E, EXXH) and B (E, QXXD). The weaker B sites in ferritin contrast with stronger B sites (E, EXXH) in diiron carboxylate oxygenases, explaining diferric oxo/hydroxo product release in ferritin vs. diiron cofactor retention in oxygenases. Codons for Q/H and D/E differ by single nucleotides, suggesting simple DNA mutations relate site B diiron substrate sites and diiron cofactor sites in proteins. The smaller k(cat) values in chimeras indicate the absence of second-shell residues important for ferritin substrate-product channeling that, when identified, will outline the entire iron path from ferritin pores through the F(ox) site to the mineral cavity.
Authors:
Xiaofeng Liu; Elizabeth C Theil
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S.     Date:  2004-05-27
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  101     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2004 Jun 
Date Detail:
Created Date:  2004-06-09     Completed Date:  2004-08-17     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8557-62     Citation Subset:  IM    
Affiliation:
Center for BioIron at Children's Hospital Oakland Research Institute, 5700 Martin Luther King Way, Oakland, CA 94609, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Binding Sites
DNA / genetics
Ferritins / chemistry*,  genetics,  metabolism*
Humans
Hydrogen-Ion Concentration
Kinetics
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Peroxides / chemistry,  metabolism
Recombinant Fusion Proteins / chemistry,  genetics,  metabolism
Grant Support
ID/Acronym/Agency:
DK20251/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Peroxides; 0/Recombinant Fusion Proteins; 9007-49-2/DNA; 9007-73-2/Ferritins
Comments/Corrections

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