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Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry2006; 11(8); 1075-1086; doi: 10.1007/s00775-006-0141-6

Ferritin-catalyzed consumption of hydrogen peroxide by amine buffers causes the variable Fe2+ to O2 stoichiometry of iron deposition in horse spleen ferritin.

Abstract: Ferritin catalyzes the oxidation of Fe2+ by O2 to form a reconstituted Fe3+ oxy-hydroxide mineral core, but extensive studies have shown that the Fe2+ to O2 stoichiometry changes with experimental conditions. At Fe2+ to horse spleen ferritin (HoSF) ratios greater than 200, an upper limit of Fe2+ to O2 of 4 is typically measured, indicating O2 is reduced to 2H2O. In contrast, a lower limit of Fe2+ to O2 of approximately 2 is measured at low Fe2+ to HoSF ratios, implicating H2O2 as a product of Fe2+ deposition. Stoichiometric amounts of H2O2 have not been measured, and H2O2 is proposed to react with an unknown system component. Evidence is presented that identifies this component as amine buffers, including 3-N-morpholinopropanesulfonic acid (MOPS), which is widely used in ferritin studies. In the presence of non-amine buffers, the Fe2+ to O2 stoichiometry was approximately 4.0, but at high concentrations of amine buffers (0.10 M) the Fe2+ to O2 stoichiometry is approximately 2.5 for iron loadings of eight to 30 Fe2+ per HoSF. Decreasing the concentration of amine buffer to zero resulted in an Fe2+ to O2 stoichiometry of approximately 4. Direct evidence for amine buffer modification during Fe2+ deposition was obtained by comparing authentic and modified buffers using mass spectrometry, NMR, and thin layer chromatography. Tris(hydroxymethyl)aminomethane, MOPS, and N-methylmorpholine (a MOPS analog) were all rapidly chemically modified during Fe2+ deposition to form N-oxides. Under identical conditions no modification was detected when amine buffer, H2O2, and O2 were combined with Fe2+ or ferritin separately. Thus, a short-lived ferritin intermediate is required for buffer modification by H2O2. Variation of the Fe2+ to O2 stoichiometry versus the Fe2+ to HoSF ratio and the amine buffer concentration are consistent with buffer modification.
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Publication Date: 2006-07-29 PubMed ID: 16896807DOI: 10.1007/s00775-006-0141-6Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigated the changes in proportions of iron(II) ions and oxygen during iron deposition in horse spleen ferritin, discovering influences of amine buffers and implicating hydrogen peroxide as a product. H2O2 is proposed to react with amine buffers, and this reaction is proved by identifying buffer modification during Fe2+ deposition.

Chemical Reactions in Ferritin

  • Ferritin is a protein complex that assists in oxidation of iron(II) ions (Fe2+) to form a mineral core of iron(III) oxy-hydroxide by using oxygen.
  • The study identifies that under different conditions, the proportions of Fe2+ to oxygen changes, which could influence its biological functions.
  • When the ratio of Fe2+ to the specific ferritin found in the horse spleen (HoSF) is large, the proportion of Fe2+ to oxygen is 4, which signifies that oxygen reduces to water. However, at lower Fe2+ to HoSF ratios, the proportion drops nearly to 2, indicating hydrogen peroxide (H2O2) as a by-product.

Role of Amine Buffers

  • The study proposes an important role of certain types of chemical buffers, specifically amine buffers, in these reactions.
  • In the absence of amine buffers, the ratio of Fe2+ to oxygen remains at 4. However, in the presence of high concentrations of amine buffers, the ratio changes to approximately 2.5.
  • This suggests that H2O2 reacts with these buffers, causing the change in iron deposition stoichiometry. This hypothesis is confirmed by identifying chemical modifications in the buffers when they partake in reactions during Fe2+ deposition.

Buffer Modification Proven

  • Mass spectrometry, NMR, and thin layer chromatography were used to compare buffers before and after the reaction, providing direct evidence of chemical modifications.
  • The buffers used—Tris(hydroxymethyl)aminomethane, MOPS, and N-methylmorpholine—were all found to be chemically altered during Fe2+ deposition to form N-oxides.
  • This alteration in buffers only happened when H2O2, oxygen and Fe2+ were incorporated with ferritin, indicating a short-lived ferritin intermediate as prerequisite for this alteration.
  • These findings hint that the presence and concentration of amine buffers to some extent control the ratio of iron to oxygen during ferritin-assisted iron deposition.

Cite This Article

APA
Zhang B, Wilson PE, Watt GD. (2006). Ferritin-catalyzed consumption of hydrogen peroxide by amine buffers causes the variable Fe2+ to O2 stoichiometry of iron deposition in horse spleen ferritin. J Biol Inorg Chem, 11(8), 1075-1086. https://doi.org/10.1007/s00775-006-0141-6

Publication

Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry
ISSN: 0949-8257
NlmUniqueID: 9616326
Country: Germany
Language: English
Volume: 11
Issue: 8
Pages: 1075-1086

Researcher Affiliations

Zhang, Bo
  • Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Wilson, Phillip E
    Watt, Gerald D

      MeSH Terms

      • Amines
      • Animals
      • Buffers
      • Catalysis
      • Ferritins / chemistry
      • Horses
      • Hydrogen Peroxide / chemistry
      • Iron / chemistry
      • Oxygen / chemistry
      • Spleen / chemistry

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      Citations

      This article has been cited 1 times.
      1. Gehrer CM, Mitterstiller AM, Grubwieser P, Meyron-Holtz EG, Weiss G, Nairz M. Advances in Ferritin Physiology and Possible Implications in Bacterial Infection. Int J Mol Sci 2023 Feb 28;24(5).
        doi: 10.3390/ijms24054659pubmed: 36902088google scholar: lookup
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