Reconstitution of manganese oxide cores in horse spleen and recombinant ferritins

Fiona C. Meldrum, Trevor Douglas, Sonia Levi, Paolo Arosio, Stephen Mann

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Abstract

The formation of Mn(III) oxyhydroxide (MnOOH) cores within the nanoscale cavity of the iron storage protein ferritin has been investigated by electron microscopy and visible absorption spectroscopy. At pH 8.9, discrete amorphous MnOOH cores were formed within horse spleen apoferritin at a range of metal:protein ratios, as well as in ferritin molecules seeded with a small ferrihydrite nucleus. Analysis of the resultant core size distributions showed that the reconstitution of horse spleen apoferritin with Mn(II) was similar to that observed previously for Fe(II) reconstitution in recombinant human L-chain ferritin, suggesting that horse spleen apoferritin does not exhibit Mn(II) oxidase activity at pH 8.9. Reconstitution with MnOOH shows essentially "all-or-nothing" behavior in which many protein molecules remain unmineralized whilst others are loaded to maximum capacity. Kinetic studies showed no significant differences between horse spleen ferritin, recombinant H- and L-chain homopolymers, and H-chain variants containing site-directed modifications at the ferroxidase and putative Fe nucleation centers. Our results indicate that the reconstitution of ferritin with MnOOH cores proceeds by a nonspecific pathway. We propose that the outer surface of the protein inhibits the development of MnOOH nuclei in bulk solution whereas the inner surface is inactive, enabling nucleation and growth to proceed unperturbed within the cavity. One possibility is that differences in the general polyelectrolyte properties of these two surfaces, rather than site-specific charges, account for the "Janus" behavior of the molecule. A similar mechanism might also increase the specificity of iron oxide mineralization in ferritins that lack ferroxidase centers.

Original languageEnglish
Pages (from-to)59-68
Number of pages10
JournalJournal of Inorganic Biochemistry
Volume58
Issue number1
DOIs
Publication statusPublished - 1995

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ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

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