Elimination of N-linked glycosylation sites from the human IgA1 constant region: effects on structure and function.
IgA1 Abs possess conserved N-linked glycosylation sites in the second C region and secreted tailpiece domains. To understand the role of these carbohydrates in the structure and function of human IgA1, site-directed mutants that produce human IgA1 lacking either one or both of the N-linked carbohydrate sites have been produced. When the mutant heavy chains are expressed in myeloma lines producing the relevant kappa-light chain, efficient secretion of the monomer and dimer forms of IgA1 is seen. In addition, higher polymer forms of the IgA molecules lacking the third domain carbohydrate, either singly or in the double mutant, are present. Functional analysis of the IgA1 proteins has shown significant differences between the various mutants and wild-type IgA. The carbohydrate mutants show a reduced affinity for their target Ag, dansyl. All of the IgA1 molecules retained the ability to bind to the polymeric Ig receptor. C3 binding was observed for all of the IgA molecules, with the IgA mutants lacking the third domain carbohydrate showing a reduced ability to bind C3; however, IgA did not effectively activate the alternative pathway, as determined by factor B cleavage and terminal complex binding. These studies demonstrate that N-linked glycosylation in the constant domain of human IgA1 plays an important role in the biologic properties of IgA1.
Chuang
P D
PD
The Molecular Biology Institute, Los Angeles, CA 90095, USA.
Morrison
S L
SL
D003167:Complement Activation / Q000276:immunology; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism*; D007127:Immunoglobulin Constant Regions / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed / Q000235:genetics*; D011485:Protein Binding / Q000276:immunology; D011961:Receptors, Fc / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics / Q000276:immunology*
D007070:Immunoglobulin A; D007127:Immunoglobulin Constant Regions; D011961:Receptors, Fc