Metal ion binding to apo, holo, and reconstituted horse spleen ferritin

doi:10.1016/0162-0134(94)00050-K

Journal of Inorganic Biochemistry

Volume 59, Issue 1, July 1995, Pages 15-27

https://doi.org/10.1016/0162-0134(94)00050-K Get rights and content

Abstract

The binding of Cd²⁺, Zn²⁺, Cu²⁺, Ni²⁺, Co²⁺, Mn²⁺, and Mg²⁺ to apo, holo, reconstituted horse spleen ferritin (HoSF), and native holo HoSF with phosphate removed was measured by gel-exclusion chromatography. Three classes of strong binding interactions (K_d < 10⁻⁷ M) with apo HoSF at pH 7.5 were found for the various M²⁺ studied: high stoichiometric binding (30–54 M²⁺/HoSF) for Cd²⁺, Zn²⁺, Cu²⁺, with two protons released per metal bound; intermediate binding (16 M²⁺/HoSF) for Ni²⁺ and Co²⁺, with one proton released per metal bound; and low levels of binding (2–12 M²⁺/HoSF) for Mn²⁺, Mg²⁺, and Fe²⁺, with <0.5 protons released per metal bound. M²⁺ binding to apo HoSF was nearly abolished at pH 5.5, except for Fe²⁺ and Cu²⁺, which remained unaffected by pH alteration. Holo HoSF bound much higher levels of M²⁺, a result directly attributable to the presence of phosphate binding sites. This conclusion was confirmed by decreased binding of M²⁺ to HoSF reconstituted in the absence of phosphate and by native holo HoSF with phosphate chemically removed. The binding of Cd²⁺ to apo HoSF was 54 per HoSF, but in the presence of developing core, the amount bound decreased to about 30 Cd²⁺/HoSF. This result indicated that Cd²⁺ and developing core were competing for the same sites on the HoSF interior, suggesting that 24 of the Cd²⁺ were bound to the inside surface. No other M²⁺ studied bound to the interior of HoSF by this criterion. Several of the M²⁺ appeared to bind strongly to the phosphate-free mineral core surface in reconstituted HoSF.

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^☆: This research was supported by Research Grant 5R01 DK36799-05 from the National Institutes of Health.

¹: SP, BW, and CL were partially supported by the Undergraduate Research Program of the College of Physical and Mathematical Sciences at Brigham Young University.

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Metal ion binding to apo, holo, and reconstituted horse spleen ferritin☆

Abstract

Adv. Inorg. Chem.

J. Biol. Chem.

FEBS Lett.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Philos. Trans. R. Soc. London B

Annu. Rev. Biochem.

Eur. J. Biochem.

Annu. Rev. Biochem.

Nature

Biochem. J.