Evidence for the molten globule state of human apo-ceruloplasmin

The conformational features of copper-free ceruloplasmin (CP), as compared to the hole-protein, were evaluated utilizing far- and near-UV circular dichroism and fluorescence spectroscopy. The results obtained indicate that apo-CP maintains the secondary structure of the hole-protein, while the tertiary interactions are much weaker. In addition, the removal of copper from the hole-protein leads to the exposure of hydrophobic patches to solvent, as shown by the fact that apo-CP, at variance from the hole-protein, binds the hydrophobic probe ANS. It is proposed that the CP molecule, upon copper removal, acquires the conformational features typical of a molten globule, which might be the conformational state of CP during its biosynthesis before metal incorporation.