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Divalent cation binding to ceruloplasmin

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Abstract

Binding of calcium to human and sheep ceruloplasniin was investigated by metal substitution with manganese and competitive displacement of bound manganese by calcium monitored by electron paramagnetic resonance spectroscopy. The K d for calcium was found to be 1.4mm. Magnesium also bound to ceruloplasmin, with K d = 0.3 and 0.7 mm for the human and sheep protein, respectively. The thermal stability of ceruloplasmin, as studied by differential scanning calorimetry, was affected by calcium but not by magnesium. A considerable increase of the T m value, from 73.8 to 83.1°C, was observed for sheep ceruloplasmin in the presence of calcium. The T m value of the human protein was only slightly altered by calcium (from 85.1 to 87°C). The interaction of ceruloplasmin with the chromatographic material used for its isolation, Sepharose 4B derivatized with chloroethylamine, was weakened by calcium. This allowed us to set up a novel purification scheme that made it possible to efficiently isolate ceruloplasmin and prothrombin from plasma with the same single-step chromatography.

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Authors and Affiliations

  1. Department of Biochemical Sciences, University of Rome ‘La Sapienza’, Rome, Italy

    Giovanni Musci, Maria Carmela Bonaccorsi di Patti, Raffaele Petruzzelli, Anna Giartosio & Lilia Calabrese

  2. Department of Organic and Biological Chemistry, University of Messina, Messina, Italy

    Giovanni Musci

  3. Department of Biology, Third University of Rome, Rome, Italy

    Raffaele Petruzzelli & Lilia Calabrese

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  1. Giovanni Musci
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  2. Maria Carmela Bonaccorsi di Patti
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  3. Raffaele Petruzzelli
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  4. Anna Giartosio
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  5. Lilia Calabrese
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Musci, G., Bonaccorsi di Patti, M.C., Petruzzelli, R. et al. Divalent cation binding to ceruloplasmin. Biometals 9, 66–72 (1996). https://doi.org/10.1007/BF00188092

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  • DOI: https://doi.org/10.1007/BF00188092

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