Effects of oxidative stress on some physiochemical properties of caeruloplasmin.
We report the effects of oxidative stress generated by low-intensity u.v. irradiation (366 and 254 nm), dialysis against ascorbate and isolated stimulated neutrophils on some physicochemical properties of caeruloplasmin. Low-intensity u.v. irradiation resulted in a loss of ferroxidase activity and 610 nm absorption, changes previously reported to occur during storage and manipulation of caeruloplasmin. These alterations were found to correspond to aggregation of the protein, induction of visible fluorescence (excitation, 360 nm; emission, 454 nm), changes in c.d. spectra which were indicative of alterations in protein conformation, loss of half-cystine, tryptophan and tyrosine content and loss immunoreactivity. The changes in the far-u.v. c.d. spectrum of caeruloplasmin were more pronounced than those observed for u.v.-irradiated IgG. Similar c.d. changes and induction of fluorescence were observed following dialysis of caeruloplasmin against ascorbate or exposure to stimulated neutrophils. It is concluded that the lability of caeruloplasmin may arise from oxidative modification, in addition to the previously described susceptibility of this protein to proteolysis.
Winyard
P G
PG
Department of Rheumatology, University of Birmingham Medical School, U.K.
Hider
R C
RC
Brailsford
S
S
Drake
A F
AF
Lunec
J
J
Blake
D R
DR
D000596:Amino Acids; D002570:Ceruloplasmin; D002851:Chromatography, High Pressure Liquid; D005453:Fluorescence; D006801:Humans; D007074:Immunoglobulin G; D009504:Neutrophils; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013997:Time Factors; D014466:Ultraviolet Rays
D000596:Amino Acids; D007074:Immunoglobulin G; D002570:Ceruloplasmin