The effect of azo dyes on heat aggregation of IgG.
The mechanism of IgG heat aggregation was studied using IgG aggregates complexed with azo dyes to increase their solubility and stability. Heat dependent and heat independent steps of aggregation were differentiated. On heating IgG at the dye concentration exceeding 100 times that of protein, mainly dimers are formed, as judged from ultracentrifugation and chromatographic analysis, whereas high molecular weight derivatives appear at room temperature when the protein/dye ratio is decreased. The analysis of spectral changes following either the attachment or removal of the dye from IgG aggregates implies that only a part of the dye molecules is bound firmly and directly to the protein binding sites. These dye molecules which are easily removed by adsorption to cellulose or reduced by dithionate but migrate together with IgG aggregates on chromatography and electrophoresis, are supposed to constitute that part of the micelle which extrudes from the binding site and, hence, is fixed indirectly to protein. Various proteins with predominant beta-structure were also found to bind azo dyes when heated.
Piekarska
B
B
Institute of Medical Biochemistry, N. Copernicus Academy of Medicine, Kraków, Poland.
Konieczny
L
L
Rybarska
J
J
D001391:Azo Compounds / Q000494:pharmacology*; D004396:Coloring Agents / Q000378:metabolism; D003208:Concanavalin A / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006801:Humans; D007074:Immunoglobulin G; D011108:Polymers; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D012967:Sodium Dodecyl Sulfate; D013053:Spectrophotometry
D001391:Azo Compounds; D004396:Coloring Agents; D007074:Immunoglobulin G; D011108:Polymers; D003208:Concanavalin A; D012967:Sodium Dodecyl Sulfate