Studies of aglycosylated chimeric mouse-human IgG. Role of carbohydrate in the structure and effector functions mediated by the human IgG constant region.
Chimeric mouse-human IgG was used to study the structural and functional roles of the carbohydrate present in the CH2 domain of human IgG molecules. To remove this N-linked carbohydrate, Asn-297, the oligosaccharide attachment residue, was changed to either Gln (a conservative replacement) or His for IgG1 or Lys for IgG3 (nonconservative replacements) by site-directed mutagenesis. Carbohydrate-deficient antibodies are properly assembled and secreted and bind Ag and protein A. However, aglycosylated IgG are more sensitive to most proteases than their corresponding wild-type IgG, indicating some conformational changes have occurred. Aglycosylated IgG do not bind to the human Fc gamma RI and do not activate C; depending on the isotype, C1q binding ability is either completely lost (IgG1) or dramatically decreased (IgG3). The serum half-life in mice of aglycosylated IgG1-Gln remains the same as wild-type IgG1, 6.5 +/- 0.5 days, whereas aglycosylated IgG3-Gln has a shorter half-life, 3.5 +/- 0.2 days, compared to that of wild-type IgG3, 5.1 +/- 0.4 days. These results indicate the carbohydrate interposed between CH2 domain of human IgG is necessary to maintain the appropriate structure for the maintenance of many of the effector functions dependent on the CH2 domain.
Tao
M H
MH
Department of Microbiology, Columbia University College of Physicians and Surgeons, New York 10032.
Morrison
S L
SL
D000818:Animals; D005260:Female; D005822:Genetic Vectors; D006031:Glycosylation; D006207:Half-Life; D006801:Humans; D006868:Hydrolysis; D007127:Immunoglobulin Constant Regions / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D007074:Immunoglobulin G / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D009844:Oligosaccharides / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010447:Peptide Hydrolases; D011994:Recombinant Proteins / Q000378:metabolism / Q000502:physiology; D013329:Structure-Activity Relationship; D014162:Transfection
D007127:Immunoglobulin Constant Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D009844:Oligosaccharides; D011994:Recombinant Proteins; D010447:Peptide Hydrolases