Delineation of the amino acid residues involved in transcytosis and catabolism of mouse IgG1.
The MHC class I-related receptor, FcRn, is involved in both the transcytosis of serum gamma-globulins (IgGs) and in regulating their serum persistence. The interaction site of FcRn on the Fc region of rodent IgG has been mapped to residues at the CH2-CH3 domain interface using site-directed mutagenesis and x-ray crystallographic analyses. In the current study, the role of individual residues (H310, H433, and N434) at this interface in mediating the Fc-FcRn interaction has been investigated using recombinant, mutated Fc hinge fragments derived from mouse IgG1. In addition, two highly conserved Fc histidines (H435 and H436) have been mutated to alanine, and the resulting mutated Fc hinge fragments were analyzed in both transcytosis and pharmacokinetic studies in mice and in competition binding assays using recombinant, soluble FcRn. The analyses indicate that mutation of H310, H435, and, to a lesser extent, H436 to alanine results in reduced activity of the Fc hinge fragments in both in vivo and in vitro assays. Thus, in addition to the previously defined role of 1253 in the FcRn-IgG interaction, these histidines play a key role in mediating the functions conducted by this Fc receptor. The effects of these mutations on binding of Fc hinge fragments to staphylococcal protein A have also been analyzed and demonstrate a partial, but not complete, overlap of the FcRn and staphylococcal protein A interaction sites on mouse IgG1.
Medesan
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Department of Microbiology and Cancer Immunobiology Center, University of Texas, Southwestern Medical Center, Dallas 75235, USA.
Matesoi
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Radu
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Ghetie
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Ward
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D000596:Amino Acids / Q000032:analysis* / Q000235:genetics / Q000502:physiology*; D000818:Animals; D000831:Animals, Newborn; D001692:Biological Transport / Q000276:immunology; D005260:Female; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism; D007700:Kinetics; D008431:Maternal-Fetal Exchange / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed / Q000276:immunology; D011247:Pregnancy; D017452:Receptors, IgG / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D013205:Staphylococcal Protein A / Q000378:metabolism
D000596:Amino Acids; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D017452:Receptors, IgG; D013205:Staphylococcal Protein A