title: A point mutation in the Ch3 domain of human IgG3 inhibits antibody secretion without affecting antigen specificity.
abstract: Immunoglobulins (Ig) require correct folding and assembly of both heavy (H) and light (L) chains to form a functional H2L2 dimer that is secreted from plasma cells. This process is dependent upon the endoplasmic reticulum (ER) chaperone BiP, which targets improperly, folded or assembled Ig molecules for degradation. While investigating the mechanism of low IgG3 secretion, we identified a missense mutation L368P in the Ch3 region of the human gamma3 H-chain that was associated with impaired secretion of intact and functional Ig. The non-secreted H-chains displayed slower electrophoretic migration than secreted H-chains, consistent with them being glycosylated in the ER but not fully processed in the golgi apparatus and secretory pathway. Reversion of the mutated codon to wild type restored secretion of the IgG3, which displayed the same fine specificity for antigen as non-secreted IgG3. However, the non-secreted IgG3 was not opsonic in an in vitro phagocytosis assay. The results indicate that correct IgG3 Ch3 domain folding is essential for secretion and effective function but does not affect specificity for antigen.
authors.affiliation: The Biomedical Research Centre, University of British Columbia, 2222 Health Sciences Mall, Vancouver, BC, Canada V6T 1Z3. gmclean@brc.ubc.ca
authors.forename: Gary R
authors.initials: GR
authors.lastname: McLean
authors.forename: Marcela
authors.initials: M
authors.lastname: Torres
authors.forename: Brendon
authors.initials: B
authors.lastname: Trotter
authors.forename: Michela
authors.initials: M
authors.lastname: Noseda
authors.forename: Steve
authors.initials: S
authors.lastname: Bryson
authors.forename: Emil F
authors.initials: EF
authors.lastname: Pai
authors.forename: John W
authors.initials: JW
authors.lastname: Schrader
authors.forename: Arturo
authors.initials: A
authors.lastname: Casadevall
mesh_terms: D019943:Amino Acid Substitution; D000906:Antibodies / Q000378:metabolism*; D000941:Antigens / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D017354:Point Mutation; D017434:Protein Structure, Tertiary
chemical_list: D000906:Antibodies; D000941:Antigens; D007074:Immunoglobulin G