[Characteristics of rat ceruloplasmin from the serum of animals, which received salts of silver with food].
Abiogenic Ag(I) ions have electronic structure, similar to Cu(I) ions and can compete with Cu(I) for binding sites of proteins which transport copper from extracellular media to sites of cuproenzyme formation in the cell. Rodents receiving Ag-salts with food develop extracellular deficiency of copper associated with ceruloplasmin (Cp, the major copper-transporting protein in blood serum of vertebrates). The present work focuses on the studies of biochemical and physicochemical properties of Cp, obtained from blood serum of rats, which received AgCl with food for 4 weeks (Ag-rats). Cp-fractions from blood serum of Ag-rats (Ag-Cp) were obtained by ion-exchange chromatography with stepped gradient of NaCl. Each fraction was tested for oxidase and ferroxidase activities by direct measurement of catalytic activity in the gel, and for specific activity in holo-Cp in oxidation of chromogenic substrate. Molecular mass, electrophoretic mobility and ratio of apo- and holo-forms in Ag-Cp fractions were evaluated by immunoblotting. Ag-Cp samples did not contain products of spontaneous partial proteolytic degradation, characteristic of holo-Cp samples. Fractions of Ag-Cp and holo-Cp (from blood serum of control rats) were compared by optical spectra, tertiary structure, susceptibility to thermal denaturation, and by atomic Cu and Ag content. Ag-Cp contained 1-2% Cp, which is similar by spectral and catalytic properties with holo-Cp. [Ag]:[Cu] ration in Ag-Cp samples was about 4:1. As evidenced by circular dichroism and differential scanning calorimetric studies, the major apo-fraction of Ag-Cp lacked tertiary structure of native Cp and was significantly misfolded, which might explain its resistance to spontaneous partial proteolytic degradation.
A N
AN
Skvortsov
E Iu
EIu
Il'icheva
E A
EA
Zatulovskiĭ
A N
AN
Savel'ev
N B
NB
Tsymbalenko
M M
MM
Shavlovskiĭ
L V
LV
Puchkova
D000818:Animals; D002570:Ceruloplasmin / Q000378:metabolism*; D003300:Copper / Q000097:blood; D017136:Ion Transport / Q000187:drug effects; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D018030:Silver Compounds / Q000493:pharmacokinetics* / Q000494:pharmacology*
D018030:Silver Compounds; D003300:Copper; D002570:Ceruloplasmin