A circular dichroism study of undegraded human ceruloplasmin.
The CD spectrum of human ceruloplasmin (Cp) has been studied between pH 6.90 and 12.00 in the far-ultraviolet, near-ultraviolet, and visible light regions. The spectrum in the far-ultraviolet region showed that undegraded holo and apo single-chain ceruloplasmin and a cleaved ceruloplasmin preparation have a low content of alpha helix but a high content of beta and unordered structure. A conformational transition accompanied by a decrease in beta and an increase in unordered structure occurred at pH 11.10 for intact ceruloplasmin. This transition probably involved the ionization of buried tyrosines, as shown by the increase of a near-ultraviolet band at 250 nm. The copper atoms may contribute to the stability of the native structure since the conformational transition occurred at a low pH value (10.50) in the case of apoceruloplasmin. The apo-Cp also presented a more intense CD band at 292 nm, suggesting the presence of tryptophan(s) near the environment of copper(s) in the molecule where no tyrosine residue seems to be involved. The spectrum between 320 and 700 nm of intact and cleaved Cp was resolved into six Gaussian bands which were assigned to type-1 copper atoms. Important changes in only two of these bands upon pH increase (bands III at 541 nm and VII at 322 nm) confirmed the nonequivalence of the two blue coppers in human ceruloplasmin.
D002570:Ceruloplasmin; D002942:Circular Dichroism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D013053:Spectrophotometry
D002570:Ceruloplasmin