Metal-catalyzed oxidation of immunoglobulin G impairs Fc receptor-mediated binding to macrophages.
Enhanced oxidative stress is a feature of inflammatory and infectious conditions. Proteins may be important targets of oxidation and this may alter their function. We evaluated whether metal-catalyzed oxidation of IgG could alter its ability to bind to Fc receptors on macrophages. Human IgG incubated with an FeCl3/EDTA/ascorbate metal-catalyzed oxidation system resulted in a significant increase in carbonyl content, a measure of protein oxidation, compared to IgG treated with EDTA alone (control). Western blot analysis using an antibody to oxidized protein revealed an increase in antibody binding to both the heavy (Fc portion-containing) and light chains of IgG treated with the oxidizing system. Western blot analysis of papain-digested IgG confirmed oxidative modification of the Fc portion. Binding studies carried out with J774.16 macrophages demonstrated significantly diminished ability of the oxidized IgG to bind to macrophage Fc receptors compared to control IgG. These data demonstrate that IgG is susceptible to metal-catalyzed oxidation and that this impairs its ability to bind to macrophage Fc receptors. Oxidation of IgG might play a role in modulating immune function in infection and disorders associated with immune complex formation by diminishing IgG binding to phagocytic cells.
Margiloff
L
L
Department of Medicine, Long Island Jewish Medical Center, New Hyde Park, NY 11040, USA.
Chaplia
L
L
Chow
A
A
Singhal
P C
PC
Mattana
J
J
D001205:Ascorbic Acid / Q000737:chemistry; D015153:Blotting, Western; D002712:Chlorides; D004492:Edetic Acid / Q000737:chemistry; D005290:Ferric Compounds / Q000737:chemistry*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology*; D010084:Oxidation-Reduction; D010206:Papain / Q000378:metabolism; D011961:Receptors, Fc / Q000378:metabolism*
D002712:Chlorides; D005290:Ferric Compounds; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011961:Receptors, Fc; D004492:Edetic Acid; D010206:Papain; D001205:Ascorbic Acid; C024555:ferric chloride