title: Naturally occurring structural isomers in serum IgA1 o-glycosylation.
abstract: IgA is the most abundantly produced antibody and plays an important role in the mucosal immune system. Human IgA is represented by two isotypes, IgA1 and IgA2. The major structural difference between these two subclasses is the presence of nine potential sites of O-glycosylation in the hinge region between the first and second constant region domains of the heavy chain. Thr(225), Thr(228), Ser(230), Ser(232) and Thr(236) have been identified as the predominant sites of O-glycan attachment. The range and distribution of O-glycan chains at each site within the context of adjacent sites in this clustered region create a complex heterogeneity of surface epitopes that is incompletely defined. We previously described the analysis of IgA1 O-glycan heterogeneity by use of high resolution LC-MS and electron capture dissociation tandem MS to unambiguously localize all amino acid attachment sites in IgA1 (Ale) myeloma protein. Here, we report the identification and elucidation of IgA1 O-glycopeptide structural isomers that occur based on amino acid position of the attached glycans (positional isomers) and the structure of the O-glycan chains at individual sites (glycan isomers). These isomers are present in a model IgA1 (Mce1) myeloma protein and occur naturally in normal human serum IgA1. Variable O-glycan chains attached to Ser(230), Thr(233) or Thr(236) produce the predominant positional isomers, including O-glycans composed of a single GalNAc residue. These findings represent the first definitive identification of structural isomeric IgA1 O-glycoforms, define the single-site heterogeneity for all O-glycan sites in a single sample, and have implications for defining epitopes based on clustered O-glycan variability.
authors: UAB Biomedical FT-ICR MS Laboratory, MCLM 570, 1530 3rd Avenue South, Birmingham, AL 35294, USA.
authors: Kazuo
authors: K
authors: Takahashi
authors: Archer D
authors: AD
authors: Smith
authors: Knud
authors: K
authors: Poulsen
authors: Mogens
authors: M
authors: Kilian
authors: Bruce A
authors: BA
authors: Julian
authors: Jiri
authors: J
authors: Mestecky
authors: Jan
authors: J
authors: Novak
authors: Matthew B
authors: MB
authors: Renfrow
mesh_terms: D000595:Amino Acid Sequence; D006020:Glycopeptides / Q000097:blood* / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood* / Q000737:chemistry* / Q000378:metabolism; D007536:Isomerism; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009101:Multiple Myeloma; D009194:Myeloma Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D006020:Glycopeptides; D007070:Immunoglobulin A; D009194:Myeloma Proteins