[Oxidative iodination of rabbit IgG: localization of markers in an Fc-fragment and effects of modification].
A comparative study of rabbit IgG, both native and modified ones, designed to assess the functional activity of these proteins under oxidative iodination conditions has been carried out. Polyclonal IgG, its antigen-specific fraction and Iodogen as an oxidant were used. Polyclonal antibodies directed against the CH2 domain of IgG, protein A targeted at the CH-2-CH3 domain interface and ferritin testing the conformation of the antigen-binding Fv fragment, were applied as conformational probes for assessing the changes in the IgG conformation. By taking advantage of pepsin proteolysis of [125I]-IgG, from 80% to 92% of the label was found to be localized within the CH3 domain, thus implying the domain-selective nature of iodination, when the degree of modification was below 0.1 atom of iodine per IgG molecule. Yet, when the three above-mentioned conformational probes were used, considerable alterations in the conformation of not only the CH2 domain and CH2-CH3 domain interface, but in the Fv domain being a part of the Fab fragment, were observed. By using competitive enzyme immunoassay for the straightforward comparative evaluation of functional properties of "cold" (native) and 125I-modified IgG, the deleterious effect of the oxidant (Iodogen) rather than iodine atom substitution at the phenolic ring of Tyr residues was shown to be the major determinant of alterations in the IgG molecule.
Zhorov
O V
OV
Kirillova
N M
NM
Martsev
S P
SP
D000818:Animals; D015415:Biomarkers; D002850:Chromatography, Gel; D006868:Hydrolysis; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism*; D007454:Iodides / Q000378:metabolism*; D010084:Oxidation-Reduction; D010434:Pepsin A / Q000378:metabolism; D011817:Rabbits; D013205:Staphylococcal Protein A / Q000378:metabolism
D015415:Biomarkers; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D007454:Iodides; D013205:Staphylococcal Protein A; D010434:Pepsin A
Okislitel'noe iodirovanie IgG krolika: lokalizatsiia metki v Fc-fragmente i éffekty modifikatsii.